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Full text of "A manual of the Aspergilli"

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A MANUAL 
of the 

ASPERGILLI 



By 

CHARLES THOM 

Collaborator, Northern Regional Research Laboratory, 

Formerly Principal Mycologist, Bureau of 

Plant Industry, U. S. Department of 

Agriculture, Washington, D. C. 

and 
KENNETH B. RAPER 

Senior Microbiologist. Fermentation Division, Northern Regional 

Research Laboratory, Bureau of Agricultural and 

Industrial Chemistry, U. S. Department of 

Agriculture, Peoria, Illinois 




BALTIMORE 

THE WILLIAMS & WILKINS COMPANY 

1945 



Copyright, 1945 
The Williams & Wilkins Company 

Made in the United Stales of America 

Published May, 1945 
Reprinted January, 1951 



Composed and Printed at the 

WAVERLY PRESS, INC. 

for 

THE WILLIAMS & WILKINS COMPANY 

Baltimore, Md., U. S. A. 



CONTENTS 

Preface vii 

Part I. General Discussion 1 

Chapter I. Historical Introduction 3 

Chapter II. Classification, Generic Diagnosis, and Synonymy 6 

Chapter III. Morphology and Description 10 

Chapter IV. Cultivation and Examination 31 

Chapter V. Preservation of Cultures 50 

Chapter VI. Variation 63 

Part II. The Manual Proper 79 

Chapter VII. The Use of the Manual 81 

Chapter VIII. The Aspergillus clavatus Group 92 

Chapter IX. The Aspergillus glaucus Group 100 

Chapter X. The Aspergillus fumigatus Group 148 

Chapter XI. The Aspergillus nidulans Group 155 

Chapter XII. The Aspergillus ustus Group 171 

Chapter XIII. The Aspergillus flavipes Group 179 

Chapter XIV. The Aspergillus versicolor Group 183 

Chapter XV. The Aspergillus terreus Group 195 

Chapter XVI. The Aspergillus candidus Group 206 

Chapter XVII. The Aspergillus niger Group 214 

Chapter XVIII. The Aspergillus wentii Group 241 

Chapter XIX. The Aspergillus tamarii Group 250 

Chapter XX. The Aspergillus fiavus-oryzae Group 259 

Chapter XXI. The Aspergillus ochraceus Group 273 

Part III. Reference Material 287 

Chapter XXIT. Topical Bibliography 289 

Chapter XXIII. General Bibliography 319 

Chapter XXIV. Check List of Species and Genera 331 

Chapter XXV. Accepted Species, Varieties, and Mutations 360 

Index 363 



PREFACE 

Aspergillus as the name for a genus of molds dates back to Micheli 
(1729), but it was not until the middle of the 19th Century that the Asper- 
gilli began to be recognized as active agents in many decay processes, as 
occasional causes of human and animal disease, and as fermenting agents 
capable of producing valuable biochemical products. Various taxonomic 
efforts were made. DeBary, Fresenius, van Tieghem, and others described 
the particular species that they used. Bainier described in brief inadequate 
terms all the Aspergilli he found. Wilhelm and Wehmer reviewed the 
literature, described and figured the forms they knew; these were few in 
number, but the work was done so well that it fixed group types. In 
1926, Thorn and Churchbrought all of this material together in amonograph. 
The increased study devoted to the Aspergilli in recent years shows some 
of their groupings to be inadequate. In addition, a large amount of new 
material has accumulated. The Aspergilli have become increasingly 
important as responsible agents in a number of industrial fermentations. 
Many of them are being found capable of producing antibiotic substances 
and their possible use in this field will undoubtedly be exhaustively ex- 
plored. For these reasons, the need for a manual for those who wish to 
identify Aspergilli under observation, without regard to the historical 
aspects of the group, has become increasingly apparent. 

This book is definitely a manual, not a monograph. It is based upon 
comparative study of thousands of strains of Aspergilli in culture. Repre- 
sentative strains giving the range of morphology and biochemical activity 
in each species are maintained in the permanent collection of the Northern 
Regional Research Laboratory. Consistent efforts have been made to 
obtain the organisms actually used by authors who have put forward new 
nomenclature. The manual thus seeks to present under species names only 
living cultures known to the authors, although it seemed advisable to make 
a few additions based upon literature. The species included are arranged 
as far as possible into natural groups which bring together aggregates of 
strains or species agreeing in important morphological characters. For 
the most part, physiological or biochemical information, if available, in- 
dicates related activities within these groups. The names selected for use 
appear to be taxonomically correct. A large number of names are neces- 
sarily rejected. If known to belong to some unidentifiable member of a 
group, or believed from literature to be correctly placed there, each of 
these names is accounted for in the discussion of the group. If the in- 
formation available does not justify allocation to some species or species 

vii 



Vlll A MANUAL OF THE ASPERGILLI 

aggregate, the name will be found in the check list with any information 
at hand. Large gaps in our information about the Aspergilli still exist. 
Some of these are pointed out in the text. The great activity of the 
present day will undoubtedly render any arrangement of the Aspergilli 
obsolete sooner or later, but it is believed that the classification put forward 
here is at least temporarily practical. 

Recognizing that any species name for an Aspergillus appearing at any 
time in the literature may at some future time become important for 
some unanticipated reason, an alphabetical check list giving each of the 
names found, the author, the date, and the place of publication has been 
included. An index reference to page in the manual, or the method of 
disposal of the name, is added to bring the material to its greatest usefulness 
as a ready reference. 

Two types of bibliography are presented: a general bibliography, 
alphabetical to author's name and sub-indexed as to date of publication 
when necessary, includes authors of species and other investigators whose 
work is cited in the text. In addition, a topical bibliography is presented. 
Although incomplete, it is hoped that the latter will assist greatly in the 
search for special literature on particular subjects. Believing that the 
more recent literature on these subjects will generally be of the greatest 
interest and value, the material is presented chronologically. Duplication 
between the two bibliographies may or may not occur. 

A manual, if it is to facilitate the identification of these molds by the 
actual worker in the laboratory, must present descriptive and illustrative 
material in as simple form as seems consistent with sound scholarship. 
From our present point of view, the describer of a mold must know that 
mold in fruiting form under the microscope as known to the early my- 
cologists, and. know it also in the culture tube. It must be isolated as a 
pure culture and its life history and reactions followed out upon laboratory 
media. Its definite place in some one of the aggregate species or groups of 
Aspergilli should be thoroughly established; then, by careful study and 
comparison proper nomenclature should not prove difficult. 

This manual, then, seeks to serve two purposes: (1) to provide the worker 
encountering an Aspergillus with means for its identification, and hence to 
open to him the whole literature of the group, as well as the particular 
species; and (2) by enumerating all forms found in the literature, and 
indicating their proper allocation, to guide the user of that literature in 
the interpretation of names found in his reading but not known to him in 
nature, in culture, or in exsiccati. 

The authors acknowledge the cooperation of Dr. Johanna Westerdijk, 
Dr. F. H. van Beyma, and their colleagues at the Centraalbureau voor 
Schimmelcultures, at Baarn, Holland, in the free exchange of cultures and 



PREFACE IX 

information. Professor Ph. Biourge and Dr. Paul Simonart at Louvani 
put their entire collection at our service after preparing and demonstrating 
their interpretations in their own laboratory. Dr. Raoul Mosseray sent 
his extensive series of variants in the Aspergillus niger group as accumulated 
from the Belgian Congo. Dr. Adalbert Blochwitz made many comments 
and criticisms in his numerous letters. Professor Harold Raistrick and 
Mr. George Smith submitted all strains reaching their laboratory with 
full notes on their own interpretations. Cultures and photographs, some 
of which are used in this manual, have been furnished by Messrs. John and 
Edward Yuill. Series of cultures have been received from Drs. Marie 
B. Morrow and J. J. Taubenhaus in Texas, Drs. Roberta Ma and Y. K. 
Shih in China, Drs. G. Kita, R. Xakazawa, J. Hanzawa, and K. Oshima in 
Japan, Drs. G. R. Bisby and G. A. Ledingham in Canada, and Dr. H. Macy 
in Minneapolis, as well as individual strains from many correspondents. 
The laboratory collection owed much of its completeness to the punctilious 
workmanship and painstaking scholarship of Dr. Margaret B. Church. 

In the preparation of the manuscript outstanding contributions have 
been made by Dorothy F. Alexander who prepared the line drawings and 
assisted generously in the checking and proofreading of the textual material; 
by Mr. Roland W. Haines, Photographer of the Northern Regional Re- 
search Laboratory, who made all the color pictures, as well as many of the 
black and white photographs; and by Miss Nancy Brant who typed the 
manuscript in its final form. 

The authors are indebted to the Chas. Pfizer and Company Inc., Brooklyn 
New York, for underwriting the cost of reproducing the natural color 
photographs. 

Administratively, the preparation of this manual was made possible by 
the vision of Dr. O. E. May, Chief of the Bureau of Agricultural and 
Industrial Chemistry, Mr. H. T. Herrick, Director of the Northern Re- 
gional Research Laboratory, and Dr. Robert D. Coghill, Chief of the 
Fermentation Division of the Northern Regional Research Laboratory, 
who have developed the industrial exploration of the biochemical utili- 
zation of the fungi over many years. 

The Authors 



PART I 

GENERAL DISCUSSION 




Chapter I 
HISTORICAL INTRODUCTION 

Historically, the Aspergilli, as a part of moldiness of things, have always 
been a factor in man's environment, but for ages were brushed away as 
white, yellow, green, red, or black mold, with or without any attempt at 
interpretation. After the development of the microscope, men began to 
see structure. Micheli (1729) distinguished conidiophores and heads. 
He noted that the heads were rough, the spore chains or columns pro- 
ducing an uneven surface, hence he gave the name Aspergillus (rough 
head). He then marked with Latin phrases his sketches of differently 
colored moldy substances, for example, Aspergillus capitatus ochroleucus, 
probably some strain of Aspergillus ochraceus; Aspergillus capilulo pulla 
for a black form, etc. Other authors followed, using much the same ter- 
minology, but without illustrations definite enough to give knowledge of 
the structure of the heads. Thus, Haller, in 1742, put what appears to 
have been Sporodinia into the genus as A. ramosissimus, etc. There is 
just about enough certainty in the use of A. albus, A. niveus, A. capilulo 
pulla, A . purpureus, etc., to justify the continued use of the name Aspergillus 
after taking out of the aggregate the extraneous material thrown into it 
by the very scanty microscopic examination given by the early mycologists. 

Persoon (in the 1790's) threw the Aspergilli into his polyglot concept, 
Monilia, based upon the production of spores in chains resembling strings 
of beads. He made no record concerning their origin. Then Link in 
1809 went back to Micheli and based his rejection of Monilia upon the 
specification that these chains of spores must have their origin in a "head" 
(capitulum) 

Link failed to examine that head closely enough to keep out questionable 
forms although we know that in describing A. glaucus he had under his 
microscope one of that group as it was found, then and now, upon partly 
dried herbarium specimens. Correct interpretation of the structure of 
this head appears first in the work of Corda who began, about 1828, to 
publish his studies of fresh material, as seen under his microscope. Up 
to about 1850 each worker was prone to look at his predecessor's descrip- 
tions and figures, and either assign whatever he had to another man's 
species, or conclude that each specimen he had was new and add another 
group of names. Montagne complained (1856) that none of the descrip- 
tions written before Corda were identifiable, while some of us are equally 
uncertain of our ability to interpretMontagne. 

3 



4 A MANUAL OF THE ASPERGILLI 

DeBary's laboratory in the early 1850's seems to have introduced suffi- 
cient culture of the molds found to form the beginning of a permanent 
literature. This started with the recognition that the yellow perithecia, 
called Eurotium herbariorum by Link, which developed among the heads 
of Aspergillus glaucus upon his herbarium specimens, were actually borne 
upon the same mycelium (fig. 7). Fresenius, Cramer, Wilhelm, and Brefeld 
in Germany followed. Raulin and van Tieghem in France developed the 
fermentation of the tannins in gall nuts to gallic acid in the 1860's with 
comparative study of other molds as a corollary. In 1880, in Paris, 
Bainier began publishing his studies of molds as they appeared in phar- 
maceutical products. He was followed by Gueguen, the Sartorys, and others 
in France, and somewhat later by Biourge in Louvain, Belgium. 

Wehmer, in Hanover, began publishing his biochemical studies in 1891, 
which led him to develop his more pretentious monograph published 
in 1901. Blochwitz undertook to develop his "system" early in the new 
century, but the World War delayed its publication until 1929. Meanwhile, 
Thorn and Church, beginning about 1910, had published The Aspergilli 
as a taxonomic monograph in 1926. Aspergilli were listed in cryptogamic 
floras, lists, manuals, and special papers of many kinds over the whole period, 
but critical discussions were few. 

In somewhat over 200 years, an enormous mass of Aspergillus literature 
has accumulated. Justice to the writers at each stage in the development 
of our information calls for an analysis of the conditions which surrounded 
its development. Practically all of the early literature was microscopical: 
the worker confined his study to specimens brought in from natural sources, 
each of which was often assumed to be typical of some species. Each 
worker used the microscope that he had at hand and the technique of 
study already known to him. Life histories and comparative examination 
of material from many sources were disregarded. Publications appeared 
as parts of floristic studies of particular regions, as reports of organisms 
found in particular lesions of man or animals, or as observed in special 
industrial connections. After DeBary's group began to study organisms 
in comparative culture, the number of publications began to increase 
rapidly. By 1929-1930 Tamiya and Morita were able to cite 2,424 titles of 
papers which, in some way, concerned the Aspergilli, in their published 
Bibliographie von Aspergillus, 1729 bis 1928. A mathematical analysis 
of this literature was published by Tamiya in 1931. Referring to his 
table 1, 71 titles appeared in the 125 years before DeBary's 1854 paper; 
73 appeared in the next 18 years preceding Brefeld 's 1872 papers; 236 in 
the next 19 years just preceding Wehmer's oxalic acid reports in 1891. 
All of this may be called the period of physiological morphology. The 
remaining two thousand, published between 1891 and 1928, represent the 



HISTORICAL INTRODUCTION 



pure culture period. This may equally well be called the biochemical 

period. 

The taxonomic part of this literature was scattered through several 
languages and represented many schools of nomenclatorial thought. The 
man who had seen only three or four Aspergilli found no difficulty in sep- 
arating them. Each used his own descriptive terms— adequate for his 
purpose but useless to the next man with different species. Saccardo 
just published them all. Critical analyses were not available. 

In The Aspergilli (1926) as a monograph, Thorn and Church sought to 
bring together all of this taxonomic literature, as published before that 
date, and to present a critical opinion as to the proper relationship of the 
species described, whether retained in the genus or placed elsewhere. 
Some 350 names were thus accounted for, but the actual number of species 
accepted as known in culture or probably determinable from existing 
literature was given as 69 (p. 252). These were more or less arbitrarily 
considered in 11 groups. In undertaking to account for all the described 
forms, it was deemed advisable to include, in the various groups discussed, 
many forms whose published descriptions were inadequate for positive 
identification, but complete enough to indicate their affinities with known 
sections of the genus. Citation of these species in the older literature might, 
therefore, be traced to group relationship, and in that way, correlated with 
more recent studies of the same or related organisms. In addition, certain 
names were listed as entirely unidentifiable and certain other forms as 
belonging to other genera. 

Various other proposals for this purpose have been made. Blochwitz 
in 1929 published his long-delayed "System und Phylogenie," with inter- 
pretations and proposals for grouping quite different from those of Thorn 
and Church. Neill (1939) reduced the species recognized to the larger 
aggregates, paying little attention to details of head and spore formation. 
George Smith (1938), seeking industrial utility, simplified his descriptions 
and introduced many photomicrographs. He discarded the literature 
for the most part and undertook to guide the worker to the larger groups 
which could be located principally by color and shape of head, as shown by 
his figures. Dodge (1935) keyed all species whose names appear in medical 
literature, from their descriptions but without studying them in culture. 
In 1939, Biourge prepared a manuscript analysis of the genus for the 
Third International Microbiological Congress in New York. His associate, 
Dr. Simonart, came to represent him, but left because of the war. The 
paper was not presented but was transmitted to us because return to the 
author was impossible. Biourge died somewhat later. His scheme of 
classification prepared in his last years is not presented because it contains 
many things too bizarre to do justice to a man who for many years was a 
master workman, as well as a valued friend. 



Chapter II 

CLASSIFICATION, GENERIC DIAGNOSIS, AND 

SYNONYMY 

Class: Ascomycetes 

Order: Plectascineae 

Family: Aspergillaceae 
Genus: Aspergillus 

Class: Fungi Imperfecti 

Subclass: Hyphomycetes 
Order: Mucedineae 

Family: Mucedinaceae 

Subfamily : Aspergilleae 
Genus: Aspergillus 

The above classification follows Engler and Prantl. Changes in the 
names of class, order, and family appear in various proposals without 
essential differences in placement. G. W. Martin would replace the names 
Plectascineae with Eurotiales, Aspergillaceae with Eurotiaceae, and Asper- 
gillus with Eurotium in the plea that the first name applied to the ascosporic 
form determines the generic usage. Since the group has too many common 
characters to be split to advantage, and since the non-ascosporic forms 
vastly outnumber the ascosporic, it is better to forget Eurotium along 
with the technicality. In this arrangement, the name Aspergillus appears 
in its proper place among the ascosporic fungi. It also appears among the 
Hyphomycetes properly keyed to facilitate the identification of organisms 
obviously related but which do not produce ascospores as far as known. 

GENERIC DIAGNOSIS 

< 

There is progressive need for broadening the application of the name 
Aspergillus to include organisms whose structures, as determined in 
culture by microscopic study, point to membership in specific natural 
groups. There is need for analysis of the question whether the whole 
group shall be retained as Aspergillus or further divided into more closely 
related entities, such as Eurotium of Link, Aspergillopsis of Spegazzini, 
Diplostephanusoi Langeron, Sterigmatocystis of Cramer, and perhaps others. 
There are so many arguments for keeping them in a single group that the 
characterization of the genus Aspergillus used by Thorn and Church in 

6 



CLASSIFICATION, GENERIC DIAGNOSIS, AND SYNONYMY 7 

1926 has been emended and introduced here. This is followed by brief 
considerations of the other more significant synonyms. 

Aspergillus Micheli, in Nova Plantarum Genera, p. 212, Plate 91. 1729. 

Compare Link, in Obs. p. 16. 1809; Corda, in Icones Fungorum 

4:31, Tab. VII, fig. 94. 1840; and Thorn and Church, in 

The Aspergilli, p. 4. 1926. 

Vegetative mycelium consisting of septate branching hyphae, colorless, 
bright colored, or in a few forms slowly becoming brown in localized 
submerged areas, or producing brown crusts, or sclerotia; conidial apparatus 
developed as conidiophores and heads from specialized, enlarged, thick- 
walled hyphal cells (the foot-cells) producing conidiophores (stalks) as 
branches approximately perpendicular to the long axis of the foot-cell 
and usually to the surface of the substrata in or upon which they are borne ; 
conidiophores unseptate or septate, usually enlarging upward and broaden- 
ing into elliptical, hemispherical, or globose fertile vesicles bearing fertile 
cells or sterigmata either parallel and clustered in terminal groups, or 
radiating from the entire surface; sterigmata either in one series only, or 
as a primary series, each bearing a cluster of two to several secondary 
sterigmata at the apex; conidia varying greatly in color, size, shape, and 
markings, successively cut off from the tips of the sterigmata by crosswalls 
(not produced by budding), and forming unbranched chains arranged into 
radiate (globose) heads or packed into columnar masses; perithecia found 
in certain groups only, unknown in most species, cleistocarpic, thin-walled, 
producing asci and ascospores within a few weeks; sclerotia regularly found 
in some strains, occasionally found in other strains, and not found in other 
and closely related strains, mostly globose or subglobose, composed of 
polyhedral thick-walled cells. 

Eurotium Link, in Obs. p. 31, Taf. 2, fig. 44. 1809. 

Synonym : Mucor herbariorum Wiggers, in Primitiae Florae Holasticae 
as No. 1158. 1780. See also DeBary, in Bot. Ztg. 12: 425. 
1854. 

The yellow perithecia suspended in networks of hyphae above or at the 
surface of his badly dried herbarium specimens were taken by Wiggers 
(1780) as the basis of Mucor herbariorum. Link (1809) recognized the 
bodies as ascosporic, hence segregated them under the generic name 
Eurotium. Then in 1854 DeBary published proof that these perithecia 
were borne upon the same mycelium as the asexual A. glaucus fruits among 
which they developed. He then called each of his Aspergilli, Eurotium 
Aspergillus followed by the specific name, whether ascosporic strains were 
known or not. In spite of technicalities invoked by some to bolster the 



8 A MANUAL OF THE ASPERGILLI 

use of the name Eurotium for all Aspergilli, or failing in that, for all ascosporic 
strains, most workers have accepted the numerical predominance of the 
non-ascosporic strains as ample reason for the general use of the name 
Aspergillus. For practical purposes, Eurotium is not used here. 

Sterigmatocystis Cramer, in Vrtljschr. Naturf. Gesell. Zurich Jahrg. 4, 
Heft 4, p. 325, Taf. II, figs. 1-15. 1859. * 

Cramer, in 1859, published his study of a black Aspergillus from the 
human ear. Since the fruiting head differed from that of Fresenius' 
A. fumigatus by showing a primary series of sterigmatic cells radiating 
from the vesicle, each bearing a crown of several sterigmata, which in 
turn each bore a chain of spores, he made this character the basis of his 
new genus Sterigmatocystis. Cramer's name has been accepted by many 
workers, but was rejected by Wehmer, Thorn, and others on the proof that 
such use would separate strains obviously related in such a group as 
Aspergillus flavus and its allies, and even among the black Aspergilli 
studied by Cramer himself. The additional name serves no useful purpose 
as an aid to identification; hence is not recognized here. 

Euaspergillus Ludwig, in Lehrbuch des niederen Kryptogamen p. 258. 

1892. 

The proposal to apply a separate generic designation to all Aspergilli 
producing sclerotia would take out the groups typified by A. candidus, 
A. niger, A. wentii, A. tamarii, A. flavus, and A. ochraceus. No one has 
followed Ludwig. 

Aspergillopsis Spegazzini, in An. Mus. Nat. Buenos Aires Ser. 3, 13: 

434. 1911. 

The black-spored Aspergilli were described as dematiaceous, hence 
separated from all the other groups. No practical reason for accepting 
this proposal has been offered. 

Diplostephanus Langeron, in Compt. Rend. Soc. Biol. Paris 87: 343-345. 

1922. 

Under this proposal ascosporic Aspergilli with the double series of 
sterigmata would be separated with A. nididans Eidam as type. No 
technical application of nomenclatorial rules justifies the complications 
introduced. 

In addition to the above names proposed to cover blocks of species 
with particular characters in common, a series of names have been used 



CLASSIFICATION, GENERIC DIAGNOSIS, AND SYNONYMY 9 

by various authors for individual species: Alliospora for a black form; 
Ascophora nigrans for A. niger; Aspergillopsis Sopp for an unidentified 
organism; Cladosarum for Yuill's mutant of A. niger; Dimargaris for some 
white forms; Emericella and Inzengaea for A. variecolor; Mucor as a place to 
assign A. herbariorum; Sartorya for a possible ascosporic A. fumigatus. 
These names are cited in the check list but contribute nothing to this 
study of the group as a whole. 




Chapter III 
MORPHOLOGY AND DESCRIPTION 

INTERPRETATION OF PUBLISHED DESCRIPTIONS 

Basic Assumptions: In interpreting the descriptions of Aspergilli in a 
literature covering a long period of time, certain assumptions, although 
not always justified, form a working hypothesis for presumptive identi- 
fication. Conidiophore walls and conidial walls are assumed to be color- 
less and smooth, unless color or markings are either figured or described. 
Perithecia and sclerotia are assumed to be lacking unless the presence of 
such structures is specifically noted. Colors are assumed to apply to the 
general color scheme of the colony, unless specifically applied to the 
conidia, ascospores, or other details by the describer. Whereas colony 
coloration may arise from an admixture of conidial structures and varying 
amounts of vegetative hyphae, colored or uncolored, together with perithecia 
or sclerotia in greater or lesser numbers, it is assumed to result from the 
massing of conidial heads unless otherwise stated. 

Difficulties encountered in interpreting descriptions based upon color 
are less for the worker with a growing culture before him than for the one 
handling descriptive literature alone, since the presence of white, green, 
yellow-green, brown, or black heads is readily distinguished with a handlens, 
even though sparingly produced upon a colony in which another color 
predominates as in many members of the A. glaucus group, or in A.fiavipcs. 
The color of the conidial heads is often made the primary basis of species 
description. 

Extent of Study: Interpretation of descriptive literature accompanied 
and supplemented rather than preceded the study of great numbers of 
cultures so that the groups established are based upon the actual handling 
of thousands of cultures representing hundreds of forms of Aspergillus 
handled during a period of more than thirty years. Many of these were 
studied on natural substrata before their isolation. In addition, exami- 
nation of exsiccati from several large herbaria, while more or less unsatisfac- 
tory as to detail in identification of species, furnish confirmatory evidence 
of the soundness of the groupings proposed. 

Types: For a few of the specific names in use today, the type strain has 
been definitely maintained in culture. For most series, selection of a 
morphological entity to give a concrete concept back of the use of a name 
becomes a matter of critical judgment. For the purposes of this manual, 
an attempt has been made to base the use of the individual name upon the 

10 



MORPHOLOGY AND DESCRIPTION 11 

morphological picture most frequently encountered, rather than upon a 
selected strain assumed to be, but not known to be, the one first described. 
Descriptive terms: For purposes of description, a standardized use of 
terms has been adopted. Great diversity is encountered in the literature 
in various languages. Even translated into Latin, Saccardo never homo- 
genized the terms so that succeeding descriptions upon the same page use 
descriptive terms in the same sense. To make comparison with existing 
literature more convenient, the usages defined in the following pages 
will cover the morphology as definitely as possible and indicate the usages 
found in the older literature. To serve as a basis for the collection, 
interpretation, and presentation of pertinent information regarding 
Aspergilli to be studied, a guide sheet of the type used by the authors is 
presented in the introductory portion of the manual proper (p. 82). 

THE ASPERGILLUS COLONY 

Since few of the Aspergilli regularly produce perithecia and ascospores, 
a basis for identifying the majority of the molds of this group as they are 
actually encountered in nature and in culture must be found in the de- 
scription of the colonies and in the details of morphology found in the spore- 
bearing structures available. 

The vegetative mass of most Aspergilli consists of submerged mycelium 
from which only fruiting hyphae rise above the surface. Such colonies 
suggest a field of ripening grain, in which conidiophores and ripening heads 
predominate, and have been described as velvety (the German term used 
is rase) from their appearance in many species. Some species produce a 
more or less aerial felt (floccosity) of branching and interlacing hyphae 
bearing conidophores. This is characteristic of certain strains of the A. 
versicolor and A. fumigatus groups upon Czapek's solution agar and other 
culture media commonly employed, and it is normally one of the most 
striking characters of A. wentii under laboratory cultivation (fig. 1 B). 
Many strains of the A . glaucus group form long streamers of hyphae hanging 
from meat stored in cool, damp rooms and certain of these retain this 
character in laboratory culture. The character of the surface growth is 
a diagnostic characteristic which is usually fairly reliable under reasonably 
uniform conditions of culture. 

In describing the Aspergillus colony, cognizance should be taken of such 
factors as age, rate of growth, temperature of incubation, and the composi- 
tion of the substratum. Provided with this information, subsequent 
investigators can intelligently interpret their cultures in terms of species 
previously described. 

Many species and strains of Aspergillus often produce conspicuously 
zonate colonies. Most commonly, these take the form of fairly regular 




Fig. 1. Colony types in the Aspergilli. A, Aspergillus parasiticus NRRL No. 
465, heavy sporing, non-floccose colony on Czapek's solution agar, room temperature, 
10 days, X 2. B, Aspergillus wentii NRRL No. 375, light sporing, floccose colony 
grown under the same conditions, X 2. C, Aspergillus variecolor NRRL No. 1954 
characterized by the production of abundant, large perithecia, X 3. D, Aspergillus 
alliaceus NRRL No. 315 characterized by abundant black sclerotia, X 2. E, Asper- 
gillus ochraceus NRRL No. 408 on Czapek's solution agar, showing heavy sporing, 
essentially azonate colony, X 2. F, The same on hay infusion agar, strongly zonate, 
X2. 

12 



MORPHOLOGY AND DESCRIPTION 13 

concentric zones and result from an increased production of conidial 
structures periodically during the development of the colony (fig. 1 F). 
The exact conditions and factors responsible for their appearance has not 
been determined, but they obviously develop, in some way, as a response 
of the fungus to its environment. Particular strains cannot generally be 
described as either zonate or azonate, since the same organism exhibits 
both characters at different times and under different conditions. In 
contrast to concentric zones, certain species in laboratory culture char- 
acteristically develop conidial heads in localized areas only. This is 
particularly true of members of the A. glaucus group, and the A. sulphur eus 
series, with conidial heads normally more concentrated at the margins of 
slant tube cultures. 

Coremia, in the broader sense of ropes of hyphae anastomosing and 
trailing upon or near the surface of a substratum, occur more or less 
commonly in Aspergilli of certain groups; but as specialized erect aggrega- 
tions of conidiophores (StilbumAike) , such structures are described and 
figured only for A. vitelline/, of Ridley. Since Ridley apparently de- 
scribed his form only as collected upon a natural substratum, the actual 
development of a specialized structure in this form remains doubtful. 
Ridley's figure could be repeated many times by roughly drawing masses 
of conidiophores bursting through the otherwise unbroken surface of a 
rich nutrient substratum, such as seeds of cereals, producing a dense 
cluster of conidiophores. 

COLOR 

The most striking character of an Aspergillus colony is usually its color 
production. This takes two general forms: (1) color in the aerial parts, 
including hyphae, conidiophores, heads, and conidia; (2) colors appearing 
in the substratum, and representing the specific response and effect of the 
organism upon particular media. The former is universally used in the 
characterization of species, while the latter usually furnishes additional 
pertinent data. In our study of the Aspergilli, citations of color have been 
made according to the terminology employed by Ridgway since the 
publication of his "Color Standard and Nomenclature" in 1912. This 
terminology is used consistently in the present manual. 

Group Color 

There is a characteristic range of colors for each group (or collective 
species) of Aspergilli, with a much narrower range in successive cultures 
of the particular species or strains. The coloring substance may be de- 
posited in the conidia only, as in A.flavus; in the conidial wall, and more or 
less present in the sterigmata, vesicle, and upper part of the conidiophore 



14 A MANUAL OF THE ASPERGILLI 

as in A. niger; or the heads may be uncolored or nearly so, while the outer 
layers of the conidiophore wall may be colored as in A. flavipes and A. 
ochraceus. In some species of the A. glaucus group, the colony color is 
at first green with the development of conidia, then predominantly yellow 
to ferrugineous from ripening penthecia. Again, in other species of the 
same group, the walls of the conidiophores and, more particularly, aerial 
hyphae become encrusted with granules that are characteristically yellow 
in the young colony, but become reddish or ferrugineous in age. Such 
colonies are at first predominantly yellow with green heads inconspicuous 
on a yellow background and latei become rusty red or brown. 

In the A.flavus group, Saito (ly07) followed by Thorn and Church (1921, 
p. 115) found that cultures with brighter shades of green when subjected 
to a vapor of ammonia would lose the green color and assume the somber 
yellow shades of variant members of the same group, and that this reaction 
was reversible, since the vapor of acetic acid would restore or even intensify 
an original green shade. The experiments pointed to the hypothesis that 
the wide range of shades produced by mixtures of yellow and green in the 
A. flavus-oryzae group' may be attributed to racial limitations, strain by 
strain, in the range of hydrogen-ion concentration produced by metabolism. 
When a carbohydrate fermentable by the particular species is present, an 
acid reaction is promptly produced in the growing colony; as growth pro- 
gresses alkaline products are also produced. The colony color in this series, 
therefore, reflects first the intensity of the initial acidity as shown by the 
intensity of the green color reached. The persistence of this shade or its 
subsequent reduction or entire disappearance to leave a somber yellow or 
finally brown colony, represents the balancing of the two activities. As 
a result, certain strains of this group, if grown on Czapek's solution agar, 
are quickly and very persistently deep green, others become particular 
shades of green, which fade to yellow and finally some of them to brown, 
and a few forms produce no true green color but assume a somber yellow 
with the first development of conidia. 

Color changes in the conidia have not been satisfactorily worked out. 
In the A. niger group, the shade of yellow to purple-brown or black seems 
to be a strain or race character little influenced by handling which is not 
destructive of the racial entity. Each race seems to reach a fixed quanti- 
tative limit in the secretion of the coloring substance, thus reducing the 
most conspicuous diagnostic character among closely related forms to a 
quantitative rather than a qualitative basis of separation. 

Color in Conidial Walls 

The conidial walls may be smooth but carry sufficient coloring matter 
in diffused form to give the characteristic colony color. Within such 



MORPHOLOGY AND DESCRIPTION 15 

series as .4. fumigatus, A. nidulans, and A. ochraceus, however, strains or 
races may be found which vary from conidial walls smooth or nearly so, 
to walls bearing echinulations or even traceries apparently produced by 
aggregation of color substance into spinules, or bars between the outer 
and inner walls of the cells. Although smoothness and echinulation have 
received much weight in descriptive literature, observations such as the 
above would indicate that this character should only be used to separate 
nearly related strains in the same group, rather than as a group character. 
Literature on the nature of color in Aspergilli seems to begin with 
Linossier's study of aspergilline as produced by A. niger in 1891; this was 
followed more recently by Quilico, and Quilico and Di Capua in 1933. 
Disregarding the record of observations only, more pretentious work 
appeared when Bainier and Sartory undertook to use color production to 
separate members of the A. glaucus group about 1910 to 1912. Their 
experiments supplemented observations of colonies checked against a 
color chart, with a routine series of solubility and precipitation tests. 
Blochwitz (1929-1935) followed by using a series of routine solubility tests 
against all species producing bright colors but failed to coordinate his 
tests to show the relation of test to culture medium and conditions and to 
age of the culture studied. Later Gould and Raistrick (1934) and Raistrick, 
Robinson, and Todd (1937) studying the A. glaucus group, extracted and 
defined the colors found, but again failed to follow the transformations 
in the color of the particular species during the course of colony development. 
Until someone correlates color determination and composition more 
closely, color observations will continue to be useful accessory data which 
must be related to the age of the colony and to the composition of the 
medium as closely as possible to have value. 

Colors in the Substratum 

Production of bright colors in the substratum is frequent among the 
Aspergilli. The color produced by any species or race in any medium is 
dependent first on the ability of the mold to elaborate the particular 
product, and second on the presence of the necessary building material 
in the substratum. A mold may grow well upon a particular medium 
without discoloring it; a transfer from this colony to another substratum 
may turn the second medium red or yellow. 

Color in the substratum is the result of the particular Aspergillus acting 
upon the particular medium under a certain range of temperature. Most 
of the species of Aspergillus, if they produce any color, produce from a 
trace to abundant yellow in the early stages. This may persist, or give 
place to shades of orange, red, or purple. In some cases, the color fades 
out as the colony becomes older. In descriptive work, progressive changes 



16 A MANUAL OF THE ASPERGILLI 

in intensity of color, or the presence or absence of color in different sub- 
strata, make the use of closely defined shades or intensities of color in the 
substratum an unreliable means of characterizing cultures. 

Observations of colors produced in the substratum remain, however, 
very conspicuous and exceedingly useful accessory characters which aid in 
the placing of species. The describer must bear in mind that such color 
reactions are confirmative, not absolute characters in separating species. 

The final difficulty in dealing with color as a separating character rests 
in the loose use of color names, which is only partially corrected by the 
use of color standards. Comparison of the same culture by different 
individuals introduces very considerable discrepancies which become 
serious when a specific descriptive name or number from one of these 
standards is introduced into a technical description. In general, a series 
of observations giving a range of colors for a species is less liable to introduce 
errors of subsequent identification. 

MORPHOLOGY 

From the time of Micheli, the name Aspergillus (literally, rough head) 
has been used for molds with a conidiophore or stalk and spore-bearing 
head (capitulum). 

The Head 

The first structure observed in a detailed study of the colony is the 
spore-bearing head. The color, shape, size, and arrangement of such 
heads are characteristic of the species and to a lesser extent of the groups 
to which they belong (figs. 4 and 5). Wehmer (1901) roughly grouped 
his Aspergilli into Microaspergilli and Macroaspergilli on the basis of 
the size of the fruiting parts. Thus, A. fumigatus, A. nidulans, A. sydowi, 
and A. versicolor would represent Microaspergilli, while A. niger, A. 
clavatus, A. ochraceus, A. wentii, and A. tamarii would be readily classed 
as Macroaspergilli. The distinction breaks down when great numbers 
of forms are studied, but the comparative size of heads and conidiophores 
remains a useful adjunct in description. The heads in certain species 
show a consistent range of measurements and form, as in A. fumigatus 
and A. nidulans which have heads of small diameter forming columnar 
masses (fig. 4). Similarly, characteristic heads of A. niger, A. ochraceus, 
or A. wentii are globose and large (fig. 5). In other species, notably in A. 
flavus or A. candidus (fig. 60), several sizes and shapes of heads are regularly 
found in the same colony. The range of size and shape, however, remains 
characteristic. The observation of many heads in the colony, and pref- 
erably in many separate cultures, forms a better basis for description of 
sizes and measurements than limited observation. Further description 




Platk I 

A-D, Aspergillus nidulans (Eidam) Wint., XRRL No. 193: .4 (upper left), portion of colony showing 
developing peritheeia and abundant conidial heads; B (upper right), conidial heads showing typical color 
and columnar form, X 60; C (center left), photomicrograph of conidial heads showing brown-walled conidio- 
phores and green conidia, X 500; D (center right), ascospores, red in color and showing two equatorial ridges 
when seen in profile, X 1200. E and F, Aspergillus janus Raper and Thorn, NRRL No. 1787: E (lower left), 
single colony showing crowded short -stalked green heads in central area and long-stalked white heads in 
localized marginal areas, the yellow-white, floccose areas being composed largely of hiille cells; F (Jower right >, 
portion of same colony somewhat enlarged, X 8. (Color photographs by Haines, Northern Regional Research 
Laboratory. Reproduced through co-operation of Chas. Pfizer & Co., Inc.) 



MORPHOLOGY AND DESCRIPTION 17 

of the structure of the head is given in the more logical order, following 
the discussion of the stalk or conidiophore. 

The Foot-Cell 

The first step toward conidium formation in the Aspergilli is the differ- 
entiation of certain cells (the foot-cells) in the mycelium for propagative 
purposes. These cells become larger, thick-walled, and each usually bears 
a single conidiophore as a branch, perpendicular to the long axis of the 
cell (fig. 2 A) and usually about midway between the ends of the cell. In 
age these cells frequently become fantastically curved and twisted with 
their connection to vegetative hyphae inconspicuous but usually still 
determinable. These foot-cells are commonly submerged in the sub- 
stratum, although there are a number of strains of the A. glaucus, A. 
fumigatus, A. versicolor, and especially of the A. flavus-oryzae groups in 
which the conidiophores arise in this way from aerial hyphae. In A . effusus 
the foot-cells are frequently long and several of them connected together 
to form whole hyphae bearing considerable numbers of very short conidio- 
phores (fig. 71 B 3 ), hence their differentiation from the sterile or vegetative 
cells is less easily determined. Failure to recognize the foot-cells as 
present in the Aspergilli led Ferdinandsen and Winge (1920) to describe 
S. dipus, using the foot-cell as the principal diagnostic character of the 
species. The presence of such a differentiated foot-cell is proposed as an 
arbitrary character to be used in separating certain depauperate forms of 
Aspergilli, which approach the structure and appearance of the mono verticil- 
late Penicillia (Citromyces), from the Penicillia. Organisms which lack 
the typical Aspergillus head with its conidiophore and especially its foot- 
cell may be best classified elsewhere. 

The Conidiophore or Stalk 

The erect, perpendicular branch from the foot-cell constituting the 

conidiophore usually enlarges upwards toward the apex at which it dilates 

more or less definitely to form the vesicle (fig. 2 C-E). The section of 

the conidiophores from the foot-cell to the base of the conidial head is 

measured and reported in describing species. 

The conidiophore in some groups is not only septate, but each cell is 
sufficiently distinct to justify the term articulate which is frequently 
encountered in the older descriptions. In our experience in examining 
specimens, articulate conidiophores are found only in the A. glaucus group. 
In most Aspergilli the unity of the whole conidiophore is fairly accentuated. 
Septa, if present, are thin, fragile, and inconspicuous; the whole conidiophore 
is enclosed by continuous characteristically thickened walls without 
conspicuous nodes as an evidence of septation. 



18 



A MANUAL OF THE ASPERGILLI 



Thickening of the conidiophore wall may be uniform or may be greater 
at the base, thinning to negligible toward the apex. Two general sections 




Fig. 2. Development of the conidial apparatus in Aspergillus niger. A, Foot 
cell bearing young conidiophore as a vertical branch. B, Developing conidiophore, 
X 172. C and D, Development of the vesicle by swelling of the terminal portion of 
the conidiophore, X 265. E x and E*, Vesicle in optical section and surface view 
showing early development of primary sterigmata, X 265. F , Later stage in develop- 
ment of primary sterigmata, X 265. _G, Young fruiting head showing secondary 
sterigmata bearing chains of conidia, X 265. 

based upon the character of the thickening of the conidiophore wall are 
fairly readily distinguished, although the careful use of high magnification 
is occasionally necessary to separate certain strains. In the first, the 



MORPHOLOGY AND DESCRIPTION 



19 



outer surface of the wall is free from pits, warts, or roughenings, hence is 
called smooth; its structure is difficult to differentiate; the mass of the cell- 
wall appears homogeneous or nearly so under the microscope and does 
not absorb the ordinary protoplasmic stains. In some species the inside 



* 



■s, %. ^ W g§- — - Comd,a - ■ 

III gU# 




Fig. 3. A, Aspergillus niveo-glaucus, NRRL No. 127, typical head showing only 
one series of sterigmata, X 575. B,A. versicolor, NRRL No. 239, typical head show- 
ing sterigmata in two series, X 1200. 

surface of the wall shows irregular clumps or uneven thickenings. When 
broken, many of these conidiophores show uneven or jagged ends like a 
broken glass tube; in the A. niger group the broken ends split like bundles 
of laths (fig. 64 B), giving a possible clue to the method of their formation. 
In the second section, the wall appears dotted or pitted, or as interpreted 



20 



A MANUAL OF THE ASPERGILLI 





Fig. 4. Conidial heads; group types. A, Aspergillus clavatus group: conidial 
heads of A. giganteus, NRRL No. 10, showing typical clavate form, X 15. B, Asper- 
gillus glaucus group: heads of A. niveo-glaucus, NRRL No. 127, showing character- 
istic radiate pattern, X 35. C, Aspergillus nidulans group: heads of A. nidulans 
showing typical short-columnar form, X 35 (Photograph by Edward Yuill). D, 
Aspergillus flavipes group: heads of A. flavipes, NRRL No. 1959, typically barrel- 
form, or loose columnar as shown, X 18. E, Aspergillus terreus group: A. terreus, 
NRRL No. 265, heads columnar, of uniform diameter throughout, often becoming 
quite long as shown, X 22. F, Aspergillus ustus group: A. ustus, NRRL No. 1974, 
heads typically loose and radiate as shown, under certain conditions approaching 
columnar, X 22. 




Fig. 5. Conidial heads; group types. A, Aspergillus candidus group: A. 
candidus, NRRL Xo. 308, showing characteristic mature heads of different dimen- 
sions, X 18. B and C, Aspergillus niger group: B, strain NRRL No. 67, showing 
typical mature heads, X 30; C, A. niger mut. schiemanni, heads typically globose as 
shown (Photograph by Edward Yuill), X 30. D, Aspergillus wentii group: typical 
globose heads of A. wentii, NRRL No. 397, X 18. E, Aspergillus flavus-onjzae group: 
A. flavus, NRRL No. 1957, typical mature heads, X 18. F, Aspergillus ochraceus 
group: A. ochraceus, NRRL No. 398, typical mature heads splitting into divergent 
columns of conidia, X 18. 

21 



22 A MANUAL OF THE ASPERGILLI 

with low magnification, often rough or echinulate (e.g., A. flavus-oryzae 
group). The secondary thickenings in such conidiophores appear to have 
been laid down around protoplasmic areas, which, for a time at least, 
maintain contact with the primary wall outside. The size and abundance 
of the pits in the mature conidiophore wall differ with the species, but in a 
general way correspond with the rate of withdrawal of the protoplasmic 
mass from its primitive connection with the original outer wall. In some of 
the species in both groups, warts, or superficial and usually more or less 
hemispherical concretions are found on the outer surfaces of the conidiophore 
wall, sometimes few and scattered widely, again fairly numerous, but always 
unevenly distributed (e.g., A. ochraceus group). These warts, or con- 
cretions, appear to be deposits of excreted substance, possibly due to the 
evaporation of the numerous drops or globules of liquid abundantly visible 
upon the young and growing conidiophores. 

The color of the conidiophore wall may be homogeneous, or the layers 
may differ markedly in shade. In a number of the groups the entire 
wall is hyaline. In certain other groups the outer layer is yellow as in 
A. ochraceus and A. fiavipes, or it may be some shade of green, brown, or 
avellaneous. In some species the whole wall is colored for all or part of 
its length. No explanation of these color differences is available, except 
possibly the varying concentration of aspergilline (Linossier, 1891) in the 
upper part of the conidiophores of members of the A. niger group, as well as 
in the conidial heads. 

The Vesicle 

The conidiophore is usually much larger toward the apex than at the 
point of origin. At the base of the head a further dilation occurs more or 
less abruptly to produce the vesicle (blase, of the German mycologists). 
This vesicle is globose, hemispherical, elliptical, or long clavate in various 
groups of the Aspergilli and furnishes an enlarged surface for the attachment 
of spore-bearing cells. The lumen of the vesicle is continuous with that 
of the upper part of the conidiophore; a septum near the base of the head 
is occasionally, but only rarely, seen (see Corda, A. mucoroides for descrip- 
tion), and has not been regularly found in any species. 

Sterigmata (Compare Fig. 3) 

The conidia-bearing surface, represented by the fertile area of the vesicle, 
is closely covered by the simultaneous development of a layer of cells, the 
sterigmata, each in a general way perpendicular to a point on the fertile 
surface of the vesicle. In figure 3A a single layer of such cells is shown, 
each of which produces an unbranched chain of conidia. In figure 3B 
each of the first series of cells, or primary sterigmata, bears two to several 



MORPHOLOGY AND DESCRIPTION 23 

cells, the secondary sterigmata, forming a crown, or verticil, at the apex. 
Each of the secondary sterigmata bears one chain of conidia. The mech- 
anism shown in figure 3B would produce several times as many chains of 
conidia as figure 3 A. Such a head as figure 3B would be compact, whereas 
figure 3 A would represent a loose head. 

In figure 3A the cells in the single layer, each producing a chain of spores, 
are in the strict sense the sterigmata. In figure 3B cells of the first layer, 
or primary sterigmata, produce verticils of cells, the secondary sterigmata, 
each of which is in the strict sense a sterigma. The cells are usually char- 
acteristic in size and shape for series of closely related species. Where 
there are both primary and secondary series, the primary sterigmata are 
essentially supporting cells and vary much more in size and shape than do 
the secondary sterigmata. For this reason they are more useful in species 
diagnosis than the secondary series. 

Various usages are found in the literature. The primary sterigmata 
are often called basidia. The secondary sterigmata are called phialids 
because they have somewhat the shape of the pharmacists' phial (vial). 
In translating descriptions into the Latin, Saccardo apparently followed 
the describers verbatim, hence used no consistent terminology. We 
find the primary sterigmata as sterigmata, basidia,. or pseudobasidia, and 
the secondary series as sterigmata, pseudosterigmata, ramuli ("ramulis 
sporiferis") or even rami (branches); all of these usages have been homo- 
genized here into "primary and secondary sterigmata." 

Conidium Formation 

The actual spore-producing cell, or sterigma, is definitely specialized. 
It ordinarily consists of an essentially cylindrical body, which, after 
reaching a length more or less uniform for the species, narrows into a 
spore-producing tube whose diameter is fairly uniform within the species. 
Elongation is thenceforth confined to this spore-producing tube. The 
nuclei in the sterigmata divide and one of each pair of daughter nuclei 
passes into the tube; cell division follows. Parallel with the repeated 
division of the sterigma nucleus, the tube continues to elongate rapidly, 
successively cutting off new sections and pushing the older cells outward. 
Each such chain of spores typically consists then of series of equal sections 
cut from one tube or tip of a sterigma and each carries a daughter nucleus 
derived directly from the active nucleus of the sterigmatic cell at the base 
of the chain. No further divisions occur among the cells in the chains. 
Such chains often contain several hundreds of spores, or conidia, each of 
which is theoretically at least exactly like the rest, hence fully capable of 
propagating the species (fig. 6). 



24 



A MANUAL OF THE ASPERGILLI 



The Conidium, or Spore 

The conidia are thus specialized propagative cells, asexual in origin, 
produced by a complex cellular fruiting structure. This consists (1) of a 
foot-cell connected with the vegetative mycelium and usually imbedded 
in the moist substratum, (2) of a conidiophore, or stalk, rising more or less 
vertically into the air to a distance typical of the species and enlarged at 
the apex to form the vesicle which is the central unit, and (3) of a dilated 
head consisting of one or two series of cells, the outermost of which are 
specialized for the purpose of producing chains of cells (the conidia), 
each equally capable of carrying the genetic factors necessary to propagate 
the species. 






Fig. 6. Camera lucida sketches showing progressive stages in conidium formation, 
X 700. A, Initiation of conidium formation. B, Secondary sterigma bearing a 
chain of three conidia, the outermost developing characteristic roughenings by the 
deposition of coloring matter between the outer (thin) and inner (firm) wall. C, 
Sterigma bearing a chain of conidia in which differentiation of the outermost spore 
is complete. D, A single mature conidium seen in surface view. See discussion on 
page 24. 

After the conidium is separated by a septum from the mother cell or 
sterigma, it remains so attached 1 as to draw nutrients from the parent 
cell at first, while it assumes the size and shape characteristic of the species, 
then it lays down within its original, or primary wall, a secondary cell wall 
whose color, texture, and marking are those of the species. The secondary 
wall completely separates this spore from the parent cell. (fig. 6). Exact 
uniformity is not attained. An occasional cell fails to develop; some differ- 
ences in size are usually evident; markings, while characteristic in nature 
and general pattern, are not always identical as to details. For descriptive 
purposes, size ranges are therefore more important than exact measurements 
and the nature of the markings found are more important than the relative 

1 Buller (Researches on Fungi V, Chapter II, 1933) discusses the primary septum 
as having a central pore through which connections are maintained. 



MORPHOLOGY AND DESCRIPTION 25 

number and dimensions of such. Such conidia may be very thin-walled, 
delicate and readily destroyed, or firm-walled, almost impervious to stains 
and able to retain their vitality for many years. They are extremely small, 
light, and float readily in air currents. In many species, the outer layer of 
the spore wall absorbs water slowly, hence such spores tend to float in 
currents of fluid or to develop as mycelia covering the surfaces of liquid 
media. Molds being typically aerobic, normal colonies develop only on 
the surface of the substratum where oxygen is abundant and their spores 
can be discharged directly into the air. Spores developing under submerged 
conditions in the absence of adequate oxygen produce fragmentary and 
defective mycelia only. 

The "Connective" 

Descriptive literature often cites the presence of a "connective" or 
"disjunctor", a "bridge" between conidia in the chain. This is sometimes 
present, again absent, in the same microscopic preparation, and when 
seen, it appears as a short space between spores, bridged by transparent 
cell walls. This is exactly what it is. Cells cut off from a cylindrical 
tube may swell and assume subglobose form without breaking their area 
of contact, or, in the swelling and rounding up process, they may partially 
or completely break that contact leaving the original cell wall of the tube, 
within which they developed, as a bridge across the open space (fig. 64 C). 
The critical examination of the developing cells in thousands of preparations 
have failed to justify interpretations which assume the degeneration of 
every alternate cell, or fantastic fusions in the production of conidia. The 
observation of connectives is, therefore, ordinarily worthless because 
morphologically it means nothing, and it is not justified by successive 
studies of the same species. 

Endogenous Conidia 

A spore is described as endogenous if it is formed within a tube or cell 
wall of a previously existing cell. It may be extruded through a tube. 
That tube may be used once only or many times. The critical factor 
is the formation of a cell or spore within an existing specialized spore- 
bearing organ and its extrusion from that body through a fixed tube. In 
Aspergillus, the tip or tube of the sterigma elongates, a cylindrical section 
is cut off carrying the tube wall and the septum at each end as the primary 
wall of the spore itself. Within that primary wall the spore as an entity 
rounds itself up to characteristic form, deposits or lays down its own wall 
with whatever coloration or markings may be typical of the species. The 
primary wall may remain separate and distinct and in the ripe spore be 
visible under the microscope, it may be blended with the secondary wall, 



26 



A MANUAL OF THE ASPERGILLI 



or it may not be determinable on the ripe conidium by ordinary examination. 
In the sense of the definition above, no endogenous conidia appear in 
Aspergillus. In cases, the primary walls are seen to break away if ripe 
spores are mounted in fluid, often carrying with them the granular materials 
which impart the characteristic marking to the spore, hence leaving the 
wall of an .A. nigcr spore smooth (A. luteo-niger Lutz). 



.«**. 



•> 





\ 






« 



«L 




-- 






) 



• 



£$& 



Fig. 7. The development and relationship of Aspergillus glaucus and Eurotium, 

after DeBary, 1854. 



Perilhecia 

The general morphology of the perithecium of the A. glaucus group 
(Eurotium) was described and figured by DeBary (1854, 1870), while 
that of A. nidulans was described by Eidam soon thereafter (1883). 
DeBary described the yellow to orange or ferrugineous perithecia as from 
90 to 300 m in diameter, without ostiole and without specialized appendages 
(fig. 7). These perithecia are borne, above the surface of the substratum 



MORPHOLOGY AND DESCRIPTION 27 

and are never more than loosely hung in networks of hyphae; they are 
often very abundant and dominate the color of the colony. Perithecium 
formation is favored by abundance of assimilable carbohydrate, but may 
or may not be completely suppressed by its replacement with nitrogenous 
products ; transfers from the same culture have developed colonies yellow 
from perithecia when grown on sugar solution, or dense green without 
sign of perithecia upon a peptone medium or a piece of leather. 

The perithecium of Aspergillus arises from a branch which coils in various 
manners in the different species to become the ascogone, as figured by 
Dangeard from forms reported as E. herbariorum, A. flavus, A. fumigatus, 
S. ochracea, S. nidulans, by Fraser and Chambers for A. herbariorum, and 
by Dale (1909) for A. repens. No fertilization process was demonstrated 
in those studies. Dangeard (1907) reported the cells in the fruiting 
apparatus as multinucleate in E. herbariorum, but to be mononucleate 
in the other species figured. In general, however, the development of an 
ascogone, followed by the development of the perithecial wall and accessory 
cell masses from vegetative hyphae below the ascogone, has been roughly 
described and figured for the group represented by A . glaucus and A . nidulans 
but interpretation of the other specific names used by Dangeard is doubtful, 
even for grouping, on the basis of the descriptions and figures given. 

Henrard (1934), working with 15 "species" of Aspergilli, reports that 
all of them are "sexually homothallic." He summarized the literature 
by saying that homothallism in the Aspergilli has been affirmed by Kniep 
(1928) for A. repens; by Schwartz (1928) for A. ruber, A. repens, four 
other strains of "A. glaucus", and four strains of A. nidulans; by Blochwitz 
(1932) for six strains in the A. glaucus group; and by Greene (1933) for 
A. fischeri. 

So far then, as present information goes, heterothallism cannot be assumed 
to account for the great variability of the Aspergilli. 

Perithecia are regularly found in most of the species of the A. glaucus 
group and in A. fischeri, which is closely related to A. fumigatus, and in a 
series of closely related forms in the A. nidulans group. They are not 
sporadic or dependent upon unknown conditions, but are regularly pro- 
duced in media which are adequately supplied with sugars and the salts 
in routine use. Their presence in A. wentii and A. citrisporus was asserted 
by Thaxter (personal communication) without producing either material, 
or description of the ascospores. Dangeard (1907) claimed to have found 
ascospores within the sclerotia of A. niger but failed to describe them. 
Diligent search over many years has failed thus far to confirm either 
statement. Still, it may be assumed that such sclerotia may be the 
homologue of structures which under some conditions might become peri- 
thecia. 



28 A MANUAL OF THE ASPERGILLI 

The Ascospore 

So far as fully described, the ascospore of Aspergillus follows the general 
type shown in the figures and description of DeBary. In the course of 
its development, the secondary thickening of the cell-wall develops in 
the form of two symmetrical valves suggesting the arrangement found 
in the shell of a bivalve mollusk, such as the hard clam (Venus mercenaria). 
The ripe ascospore is commonly shaped as a double convex lens with the 
valves more or less closely in contact at the edges. A series of variations 
upon this basic pattern occur and characterize particular species (figs. 27, 
34, and 43). If the exospore is smooth and the margin of the valves is 
not marked by folds or ridges, figure 27A characteristic of A. re-pens appears; 
if the exospore is rough in the absence of marginal folds, figure 27D 
characteristic of A . amstelodami results ; if the exospore is rough and the 
margin of the valves bear folds or ridges, figure 43C characteristic of A. 
rugulosus and A. jischeri is produced. An extreme development of the 
marginal folds is seen in- A. variecolor, figure 43D. 

When such an ascospore germinates (fig. 8A), the figure first shown by 
DeBary (1854) develops. Exactly the same type of germination is shown 
by A. nidulans, in which as the spore swells, the valves first separate at 
one edge, then parting completely, remain on opposite sides of the germi- 
nated spore conspicuously identifiable by the bright purple-red color of 
the valves (Thorn and Church, 1926). 

Htille Cells 

Mature perithecia were described for A. nidulans by Eidam (1883) 
but without giving attention to their origin. In his description, Eidam 
pictured a loose network of hyphae more or less completely surrounding 
the perithecium containing large numbers of "Hiille" cells, terminal or 
intercalary cells which swell and become vesiculose, elliptical or almost 
globose, then develop very heavy thick walls almost obliterating the cell 
lumen (fig. 49). These cells were later noted by Dangeard (1907) who 
designated them as chlamydospores, but failed to present evidence of 
function as propagative cells. These cells are abundant in connection 
with perithecia in all strains of the A. nidulans group, but are lacking in 
A. unguis which does not produce perithecia. Cells of the same general 
character appear in sterile masses of hypae in strains of the A. ustus and 
A. flavipes groups (fig. 49), and in some strains of the A. versicolor group, 
while thick- walled, septate hyphae at least suggestive of hiille cells appear 
in Aspergillus carneus in the A. terreus group (fig. 49F). 

In the .4 . nidulans group hiille cells, "Eidamsche blasen," or chlamydospore 
(terms found in the literature) are always produced in connection with 
perithecium formation (fig. 42). In the A. flavipes, A. ustus, and A. 



MORPHOLOGY AND DESCRIPTION* 



29 




h 



A 






^f 



9 



i 






^ 



• 



a 



b 

© 



^ 



rf 



B 

Fig 8 Spore germination. A, Germinated ascospores of Aspergillus nidulans 
X 750: a, ascospore in profile showing how the two valves comprising the spore wall 
are pushed apart as the spore content enlarges and the sporelmg develops; b, the 
same in face view. B, Germinating conidia of Aspergillus niger, X 600: a, ungermi- 
nated spore ; b , spore in early st age of germination ; c , germinated spore showing rapidly 
elongating sporeling. 



30 A MANUAL OF THE ASPERGILLI 

versicolor groups, they occur in scattered aggregates of varying conspic- 
uousness, although no perithecia have been found. Occasionally, as in one 
of the A. flavipes group, foot-cells and sterile cells, which appear to be 
conidiophores ending in points instead of heads, appear. These aborted 
conidiophores vary from nearly full length to vestigial. From the study 
of such material it is believed that Eidam's hiille cells represent aborted 
conidiophores surrounding the perithecia of A. nidulans and they may be 
indicative at least of a vestigial precursor of perithecium formation when 
they occur elsewhere. 

Hiille cells do not, however, accompany the perithecia of either the A . 
glaucus group or A. fischeri. In the former, the perithecia are smooth- 
walled and naked ; in the latter, they are surrounded by a loose network of 
sterile but unspecialized hyphae. 

In A. panamensis (Raper and Thorn, 1944) aborted conidiophores 
commonly occur, but no hiille cells have yet been observed. In A. unguis 
(Thorn and Raper, 1939), which is one of the A. nidulans group lacking 
only the ascosporic phase, the long, sterile, thick-walled "spears" suggest 
homologous structures. To summarize, hiille cells are specialized structures 
that normally occur in certain groups of the Aspergilli. They are of 
somewhat questionable origin and, in our experience, are not known to 
serve any functional purpose. Schwartz (1928) figures the hiille cells of 
A. nidulans as capable of germinating and thus acting as reproductive 
cells. Since the cells of different groups are fairly characteristic, they 
often provide valuable diagnostic group and species characters. 

Sclerotia 

Definitely hard masses with characteristic surface marking and color- 
ation, and consisting of thick-walled parenchyma-like cells occur in several 
groups of Aspergilli. Such structures have not been seen in the groups 
typified by A. clavatus, A. glaucus, A. fumigatus, A. nidulans, A. ustus, 
A. versicolor, or A. terreus. On the other hand, they develop regularly in 
certain members of the A. candidus, A. niger, A. wentii, A. tamarii, A. 
flavus-oryzae, and in the A. orchaceus groups; in other members of these 
same groups grown under similar conditions, no such structures have 
been found. In occasional strains of A. flavipes dark hyphal masses are 
seen, but these are not sufficiently compact to be considered true sclerotia. 
The development of sclerotia has not been followed in sufficient detail in 
any species to fix their genetic significance, but the specialized, character- 
istic structure of the sclerotia of many species lends color to the report of 
ascospore formation by Dangeard (1907), although no one else seems to 
have been able to verify such development. Environmental factors are 
known to influence sclerotium development, but these have not been 
carefully analyzed. 



Chapter IV 
CULTIVATION AND EXAMINATION 

An Aspergillus occurring upon a natural substratum may frequently be 
identified as to group from the original material if it belongs to one of the 
large and well-marked groups. Among members of the same group, how- 
ever, differences in the nature and composition of the substratum produce 
marked contrasts in colony appearance, quantity of growth, coloration, 
measurements of fruiting structures, and in the appearance of conidiophores 
and conidial masses. Many Aspergilli can be identified as to group from 
dried herbarium materials, but the process of drying generally changes the 
colors materially and renders the hyphal masses so fragile that the morpho- 
logical details necessary for identification inside a particular group are often 
obliterated or made difficult to interpret. Although accurate placing of 
such materials is sometimes possible, a much more satisfactory identifica- 
tion may be reached by transferring fresh material to culture media of 
known composition, followed by purification of the cultures so that they 
present single species or strains for intensive study. 

Two types of material for identification are regularly encountered by one 
undertaking a study of the Aspergilli: (1) the culture already isolated by 
another worker, and (2) the moldy substratum with its natural flora of 
micro-organisms often representing several or many species, including 
miscellaneous molds, bacteria, actinomycetes, and even protozoa and other 
forms. In either case, the final decision as to the proper name of an 
Aspergillus must usually be sought in fresh cultures made by the one re- 
sponsible for identification. 

Classification within the genus has become so dependent upon observa- 
tion in pure culture that the whole subject of laboratory cultivation, includ- 
ing favorable substrata, culture making, and culture handling needs to be 
considered. 

CULTURE MEDIA 

Pure culture upon known substrata is almost essential to the identification 
of Aspergilli. Since the morphological responses to diverse nutrients, and 
especially to the stimulus of mixtures of other molds and bacteria growing 
with any particular species, is great, the study of each strain or species in 
pure culture in media of known composition is practically necessary. 

Aside from Raulin's group in Paris, most of the early culture work with 
Aspergilli was carried through upon so-called natural media. DeBary and 

31 



32 A MANUAL OF THE ASPERGILLI 

Brefeld used decoctions of horse dung variously diluted and stiffened with 
gelatin. Much of the European work was done with brewery wort; Maze 
(personal communication, 1904) used extract of white beans; potato and 
carrot decoctions have been widely used; Bainier grew his molds and de- 
scribed many of them upon sticks of licorice root; others preferred plugs of 
potato or of carrot, string beans, etc. The primary aim was to obtain an 
optimum growth of the mold under observation, rather than to analyze its 
relation to the substratum or furnish comparative data to distinguish it 
from other members of a series. 

An optimum culture substratum for comparative study of the Aspergilli 
needs to contain the necessary chemical elements in the form of pure but 
assimilable salts, supplemented by carbohydrates of such structure as to be 
available to the largest number of species, and purchasable in pure form in 
the chemical trade. Supplementary and often very important information 
must be sought from variations in the proportions of nutrients used, or in 
the introduction of widely different substances in replacement of particular 
components of media already used. The Aspergilli as they are isolated from 
nature are not very dependent upon vitamins or growth-promoting sub- 
stances. For cultivation upon solid substrata, agar is almost universally 
employed as a gelling agent. 

When the study of a large number of molds is undertaken, comparison of 
these molds under controlled and reproducible conditions of growth become 
essential. Foremost among the conditions which must be standardized is 
the culture medium, or substratum. Various authors have proposed 
standardized and reproducible formulae which in their experience have pro- 
vided uniform cultures over long periods, hence are of value for comparative 
studies. A series of such media are presented. 

Czapek's Solution Agar 

As a routine medium for comparative work, the following formula origi- 
nally adapted from Czapek (1902, 1903) by Dox (1910) has been widely 
used. Minor variations in quantities apparently do not affect the reactions. 
Cultural information given in this manual is obtained from growth upon media 
produced by this formula unless otherwise specified. 

Czapek's solution ar;ar: 

Water 1 , 000 cc . 

NaN0 3 3.0 grams 

K 2 HP0 4 1.0 gram 

MgS0 4 -7H 2 0.5 gram 

KC1 0.5 gram 

FeS(V7H 2 0.01 gram 

Sucrose (Cube or other good commercial grade) 30.0 grams 

Agar 15.0 grams 



CULTIVATION AND EXAMINATION . 33 

To reduce caramelization the sugar is added just prior to final steri- 
lization. 

Czapek's solution agar is not offered as an optimum substratum for any- 
particular species, but as a mixture approximately neutral in reaction, 
which is readily made in any laboratory in fairly uniform manner, and which 
permits moderately vigorous growth of nearly all of the saprophytic Asper- 
gilli. The quantities of mycelium and conidia produced by many forms 
are much greater upon other media, but for comparative study, a moderate 
growth of the majority of the species is more useful than the great mass of 
mycelium ard conidia which are readily obtained by using enriched sub- 
strata. 

Since the purpose of the Czapek formula is to insure the presence of the 
chemical elements required in quantities sufficient to support good growth, 
it is frequently modified as to quantities and nutrients introduced. For 
some Aspergilli such as the A. glaucus group, the addition of 20 percent or 
even 40 percent of sucrose has proved useful. Ammonium nitrate is some- 
times substituted for sodium nitrate but with the loss of information as to 
whether the mold utilizes the ammonia or the nitrate, or both. Dextrose 
is commonly substituted for sucrose. The monobasic potassium phosphate 
prevents precipitation of certain components in sterilization, but it produces 
an acid instead of a neutral medium, hence complicates many pieces of work. 
The introduction of peptone or yeast extract increases the sporulation of 
some forms. These and other changes are made, however, by investigators 
who still refer to Czapek's solution as the basis of their work. 

Biourge, in his monograph of Penicillium (1923) and in his unpublished 
Manuscript of Aspergillus (1939), put much emphasis upon the method of 
preparing the neutral Raulin medium which he used for the growth of the 
colonies analyzed in making his species diagnoses. 

Neutral Raulin's Solution — Dierckx-Biourge 

1. Magnesium carbonate 0.40 gram 

Tartaric acid 0.71 gram 

Triturate in a mortar with a few drops of distilled water and add quickly to a 
flask of distilled water; make up to 100 ml. 

2. To a liter flask with 800 to 900 ml. distilled water add: 

Sucrose 46.60 grams 

Ammonium nitrate 2.66 grams 

Ammonium phosphate 0.40 gram 

Potassium carbonate 0.40 gram 

Ammonium sulphate 0.16 gram 

Zinc sulphate 0.04 gram 

Iron sulphate 0.04 gram 

3. Add 66 to 67 ml. of the magnesium tartrate solution (1) to the mineral salt- 
sucrose solution (2) and make up to 1,000 ml. with distilled water. 



34 A MANUAL OF THE ASPERGILLI 

In his detailed study of the A. niger group, Biourge's pupil, Mosseray 
(1934a), gives his simplified Raulin's solution as follows: 

Water, distilled 1,000 ml. 

Sucrose 50 grams 

Tartaric acid 1 0.40 gram 

Magnesium carbonate 1 0.250 gram 

Ammonium nitrate 2 0.250 gram 

Potassium carbonate 2 0.40 gram 

Ammonium phosphate (NH 4 ) 2 HP0 4 1 0.40 gram 

Ammonium sulphate 1 0.20 gram 

Iron sulphate 1 (cryst.) 0.05 gram 

Zinc sulphate 1 (cryst.) 0.05 gram 

Agar-agar 1 20.00 grams 

Sterilize 120° C. for 20 minutes. 

Steinberg's Solution 

Steinberg in the course of many years' investigation of a single strain of 
Aspergillus niger (NRRL No. 334: Thorn No. 4247) developed a basic for- 
mula for testing other phases of the nutrition of his mold. It is called the 
"dibasic optimum" solution and carries mannitol instead of sucrose, sodium 
nitrite instead of ammonium nitrate, with the addition of sufficient sulphu- 
ric acid to obtain any desired reaction. Interpolations into this solution 
offer many possibilities. 

Steinberg's "dibasic optimum" (Thorn and Steinberg, 1939) 

Water (distilled in Pyrex still) 1,000.0 grams 

d-Mannitol 50.0 grams 

i^aNO-: 2.0 grams 

K 2 HP0 4 0.35 gram 

MgS0 4 -7H 2 0.25 gram 

FeS0 4 -7H 2 0.001 gram 

ZnS0 4 -7H 2 0.00088 gram 

CuS0 4 -5H 2 0.00020 gram 

MnS0 4 -4H 2 0.00012 gram 

NaMo0 4 -2H 2 0.00005 gram 

H 2 S0 4 to pH 4. 

A number of so-called natural substrata, including malt extract and hay 
infusion agars, are very useful in the study of the Aspergilli. A great ma- 
jority of species sporulate more freely upon malt extract than upon Czapek's 
solution agar (fig. 9), and for this reason it is very useful where large quan- 
tities of spores are desired. This medium is, however, less diagnostic than 
Czapek's solution. 

1 Merck reagents. 

2 Kahlbaum reagents. 



CULTIVATION AND EXAMINATION 35 

Malt Extract Agar (Blakeslee's Formula, 1915) 

Distilled water 1 ,000 cc. 

Malt extract 20 grams 

Peptone 1 gram 

Dextrose 20 grams 

Agar 20 grams 

Add dextrose just prior to final sterilization. Conidial structures are 
generally more numerous and are often borne on shorter conidiophores, and 
there is an almost complete absence of coloration in the substratum. Ex- 
cept in a few isolated cases, coloration of the spore heads themselves is not 
materially altered. While the production of exudate in the form of drops 
is not characteristic of many of the Aspergilli, it can be generally said that 
droplet formation on malt agar is much less than upon Czapek's solution 
agar. 

Hay infusion agar is very useful in the isolation of Aspergilli from nature. 
A 1 : 10 suspension of soil in sterile water is streaked on hay infusion agar 
plates and incubated for one week to 10 days. Isolations are then made 
from individual fruiting structures with the aid of a low-power binocular. 

Hay Infusion Agar 

Distilled water 1,000 cc. 

Decomposing hay 50 grams 

Autoclave for 30 minutes at 15 pounds. Filter. 

Infusion nitrate 1 , 000 cc . 

K 2 HP0 4 2 grams 

Agar 15 grams 

Adjust pH to 6.2± 

No Aspergillus makes a luxuriant growth upon this medium, but a great 
variety of forms make a limited development. Furthermore, such fruiting 
structures as are produced are generally characteristic of the different 
species present. It thus constitutes a very favorable substratum with 
which to analyze and isolate the Aspergilli occurring in soils or other natural 
substrates. The medium is likewise useful for securing limited sporulation 
of certain forms, such as Aspergillus sparsus, which fruit very sparsely upon 
Czapek's solution agar. 

Czapek's solution agar enriched with peptone or corn steeping liquor is 
often very useful. For example, in the cultivation of members of the 
glaucus group, the addition of a limited amount of peptone greatly increases 
the production of conidia, while addition of a small amount of corn steeping 
liquor (e.g., 0.2 percent) increases the growth of most forms without mark- 
edly affecting the character of the resulting colonies. 




Fig. 9. Influence of substratum. A-C, Aspergillus variecolor, NRRL No. 212, 
growing upon Czapek's solution, malt extract, and hay infusion agars, respectively; 
10 days; room temperature. Note particularly the heavy development of perithecia 
upon malt and the very sparse development of the same upon hay infusion agar. 
D-F , Aspergillus caespitosus, NRRL No. 1929, growing upon the same media under 
similar conditions. Upon Czapek's solution agar (D) dark hiille cell masses develop 
along with a limited development of conidial structures; upon malt (E) and hay 
infusion (F) agars hiille cell masses are lacking and conidial structures are very 
abundant. 

36 



CULTIVATION AND EXAMINATION 37 

SPORULATION MEDIA 

In addition to the ordinary handling of molds in the laboratory, explora- 
tion of their biochemical potentialities often necessitates the production of 
considerable quantities of spores. Whereas some of the media listed above 
may be employed for this purpose, it is generally advisable to employ 
so-called "sporulation media." Requirements of different species and 
strains vary and one frequently has to develop special solutions to meet the 
needs of particular organisms under study. Such media may be used either 
as liquid substrata or as solutions solidified with agar. In either case the 
objective is the same: to secure the maximum production of spores with the 
production of as little vegetative mycelium as possible. 3 

A number of sporulation media for use with different molds have been 
developed by Dr. A. J. Mover. Three of these will be cited, while addi- 
tional formulae may be found in the papers published by members of the 
Fermentation Division, Northern Regional Research Laboratory. 

Sporulation medium for A. niger {Moyer, Wells, Stubbs, 
Hcrrick, and May, 1937) 

Glucose 91 .3 grams 

NH 4 N0 3 0.450 gram 

KH 2 P0 4 0.072 gram 

MgS0 4 -7H 2 0.060 gram 

Beer 60.0 ml. 

Distilled water to make 1 liter 

This solution can also be used as a solid medium by the addition of 0.5 
gm. per liter CaC0 3 and 30.0 gm. per liter agar. The above solution was 
subsequently modified as follows (Gastrock, Porges, Wells, and Moyer, 
1938): 

Glucose 50.0 grams 

(NH 4 ) 2 HP0 4 0.560 gram 

KH,P0 4 0.144 gram 

MgS0 4 -7H,0 0.120 gram 

Peptone 0.20 gram 

Beer 45.0 ml. 

Distilled water to make 1 liter 

3 For the surface inoculation of nutrient solutions in small flasks or other con- 
tainers, dry spores in quantity can be removed from agar surfaces and floated on the 
liquid surface by means of a 5 mm. loop. To inoculate from a liquid culture, tae 
usual procedure is to remove a small portion of the heavily sporing mat, transfer 
this to the solution, and dislodge the spores by vigorous agitation. Spores from 
either type of culture can be suspended in water and used as inoculum in submerged, 
or shaken, cultures. The addition of sodium lauryl sulfonate to the suspending water 
in a concentration of 1 : 10,000 aids greatly in securing uniform spore suspensions. 



38 A MANUAL OF THE ASPERGILLI 

Sporulation medium for A.flavus (Moyer, Personal communication) 

Glucose (commercial) 165.0 grams 

Bacto-Peptone 1 .0 gram 

MgSCV7H 2 0.050 gram 

KH 2 P0 4 0.060 gram 

KNOj 0.500 gram 

Fe (as tartrate) 0.040 gram 

Agar 0.25 gram 

Distilled water to make 1 liter 

The above solution can be used as a solid medium by increasing the agar 
concentration to 30.0 gm. per liter. (The small amount of agar included in 
the basic formula is added solely for the purpose of increasing viscosity.) 
Incubation should be at 22° to 24° C. This factor is critical for maximum 
spore production with the kojic acid producing strain, NRRL No. 484 
(Thorn No. 3538), for which the medium was developed. This medium can 
also be successfully employed for spore production in A . niger by incubating 
cultures at 30° C. 

Abundant sporulation of many strains and species can be secured by cul- 
tivation upon bread, whole cereal grains, or various types of milled products 
of the same. Bread, if used, should not contain proprionates or other mold 
inhibitors. The material to be inoculated should be moist but in no sense 
wet, and special precautions must be taken to insure that the grain or bread 
is properly sterilized before being used. The use of grain as a substratum 
for molds dates back to prehistoric time in the fermentation industries of 
the Orient where rice was, and still is, commonly used to produce the 
"koji," or inoculum, used in the alcoholic and soya fermentation industries 
of that area. In its classic usage, the molds cultivated were mostly mem- 
bers of the A. flavus-oryzae group, but experience has shown that this 
general type of medium can be used to advantage to secure heavy sporula- 
tion of many other forms. From such material, series of surface fermenta- 
tion flasks or other vessels can be uniformly inoculated by various means 
involving aspiration of spores, or the direct transfer of heavily spore-laden 
particles. Spore suspensions can be prepared and used for seeding sub- 
merged cultures 

TYPES OF CULTURE 

Cultures for grouping and identification of the Aspergilli should be 
grown in petri dishes. At the same time an adequate number of slanted 
tubes should be inoculated and held in reserve as an uncontaminated stock 
culture. Colonies so situated in the petri dish that they can be viewed 
directly under low magnifications with the compound microscope are neces- 
sary to supply a clear picture of the structure and course of development 
of mycelium and fruiting parts. Such colonies can be obtained by several 



CULTIVATION AND EXAMINATION 39 

procedures, some of which have been developed for special tests, or for indi- 
vidual mold problems. Practices useful in bacteriology are often applicable 
to the problems of the mycologist. Selection of a procedure which satis- 
factorily provides the information necessary to describe an Aspergillus calls 
for discussion of a series of procedures commonly in use. Only in that way 
may we show why some of these are adapted to the problems of identifying 
an Aspergillus while others fail, for specified reasons, to furnish important 
observations. 

Spot Inoculations 

Over long periods and in the hands of many investigators some type of 
mass conidial inoculum has given dependable and reproducible results. The 
most common method of transfer and, on the whole, probably the most 
satisfactory one for maintaining a strain of Aspergillus, as well as other 
molds, is the removal from the stock culture of a variable mass of conidia, 
fruiting structures, or vegetative hyphae with some sort of needle or loop 
and the transfer of this material to selected positions on fresh medium. 
Practices differ. With the Aspergilli, colonies so placed as to permit radiate 
development from the point of inoculation are most satisfactory for study. 
Where there are two or three colonies to the plate, these eventually reach 
into each other's zones of influence. They may blend and become indis- 
tinct, or inhibit each other and leave sterile bands between the colonies. 
Both types of culture furnish useful data. Usually, the line where two 
colonies approach and partially or completely inhibit each other hastens 
fruiting in the adjacent margins and permits favorable examination with the 
compound microscope. The opposite margin of the same colony, unaf- 
fected by competition, furnishes at the same time the normal and symmetri- 
cal growth which is typical of the species. The one-colony plate usually 
provides the most striking exhibit of the species, but the 2- or 3-colony plate 
is the most generally useful (fig. 10). 

Once an Aspergillus has been obtained in pure culture, the most effective 
way to insure that plates will contain 1, 2, or 3 colonies, as desired, is to 
suspend a quantity of spores in melted agar at approximately 45° C, allow 
this to solidify, and then transfer small quantities of the gelled suspension 
to the surface of plates or agar tubes in the positions where the colonies are 
desired. With the Aspergilli, as with all of the molds which produce dry 
spores, it is often very difficult to secure placement of colonies at selected 
positions, and only at such positions, without the use of some type of 
wetted inoculum. 

In the routine examination of Aspergilli, where it is not essential that the 
number of colonies be limited to 3 or less, satisfactory transfers have been 
made by using a sharp nichrome wire as an inoculating needle and selecting 



40 A MANUAL OF THE ASPERGILLI 

the inoculum from a colony in a petri dish by working under a 10 X pocket 
magnifier, or under a binocular microscope of the Greenough type carrying 
similarly magnifying lenses. Material to be removed can be exactly lo- 
cated in the parent colony. It is found possible in dealing with most 
Aspergilli (1) to remove conidia from a single head, (2) to remove one or 
more heads borne upon a single hypha at the margin of the colony, or (3) 
to select vegetative hyphal tips from a single mycelial sector. It is usually 
possible to avoid (1) heads and conidia of other species or strains, (2) foreign 
mycelia, and (3) bacterial contaminations present in the substratum. 
Purity in repeated culture over many years has been possible by this pro- 
cedure. 








h 



B 



Fig. 10. Single and three-point cultures of Aspergillus foetidus on Czapek's solu- 
tion agar, room temperature, 10 days, X i approximately. Note that spore produc- 
tion extends to the colony margins in the central triangle of 'figure B, and almost to 
the outer colony margins as well. In figure A, sporulation is limited to the central 
area only. Figure B is favorable for direct examination with the low power objective 
of the compound microscope; figure A is not. 

Dilution Cultures 

If dilution cultures are to be used, the suspension of conidia should be 
sufficiently dilute so that the individual petri dish will show not more than 
six, but preferably not over three, colonies. Such dilutions are difficult to 
gauge and often result in some plates crowded with numerous colonies while 
others contain none. The colony locations are not under control. In 
general, the dilution of spores in successive water blanks with the subse- 
quent plating of aliquots from such dilutions has not been found sat- 
isfactory. 

An alternate technique involves the introduction of one loopful of inocu- 
lating material into a tube of melted agar which is then rolled or shaken, 



CULTIVATION AND EXAMINATION 41 

and a loopful removed to a second tube, followed by the same manipulation 
a third or fourth time. The higher dilutions are then poured into petri 
dishes where colonies develop. This method has all the faults of the water 
blank dilution technique. The procedure has been widely used but has not 
been followed in our study of the Aspergilli. Blakeslee (1915) and other 
mycologists have prepared such dilutions in bottles by slowly rotating the 
bottle in a cold water or ice bath, thus allowing the agar to congeal in a thin 
layer against the glass surface. This practice has little to recommend it: 
(1) the resulting colonies could be isolated more readily from a petri dish, 
and (2) it is basically impractical to make satisfactory observations regard- 
ing the character and structure of a mold colony, or the gross features of its 
fruiting structures, when observations have to be made through the curved 
and non-uniform walls of a glass bottle. 

Smears and Streak Cultures 

The practice of smearing a suspension of mold spores over the whole plate, 
or the whole length of a slanted agar tube, results in a growth of mycelium 
which covers the entire surface and usually produces a greater mass of co- 
nidia. But individual colonies are usually unidentifiable in such preparations, 
hence they are of little value in providing those critical details of colony 
habit, coloration, and texture necessary for identification and classification. 
In general, mycelium derived from different conidia of the same strain will 
intertwine without inhibition, and even anastomose, if they come into con- 
tact in the early stages of growth; for example, before any sign of conidium 
production appears. If the spacing of the spores is great enough to permit 
the establishment of small fruiting colonies before such contact is made, the 
colonies frequently do not converge and complete coverage of the surface of 
the agar with the production of maximal quantities of conidia does not oc- 
cur. Closely-placed seeding of spores is useful to obtain the largest possible 
supply of conidia; but to study the normal characters of a species, individual 
colonies must be allowed to develop without interference by others of the 
same or different species. 

Streak cultures can be employed to advantage in freeing one mold from 
another, or from other contaminating organisms. By touching a sterile, 
moistened needle to a single conidial head or other selected fruiting surface 
of limited extent, and subsequently streaking this repeatedly across the 
surface of an agar plate, isolated colonies of the desired species can usually 
be obtained. Occasionally it is desirable to streak two plates in succession 
in order to secure a satisfactory separation of colonies. In any case the 
spore source should be selected with great care and the use of a low-power 
binocular microscope or pocket magnifier is recommended. When this 
method is employed, it is desirable to reisolate from these individual colo- 



42 A MANUAL OF THE ASPERGILLI 

nies as soon as possible after sporulation begins. For freeing one species or 
strain of Aspergillus from another, or from a contaminating Penicillium, 
streaking upon Czapek's solution agar usually gives satisfactory results. 
The method finds its greatest usefulness in separating comparatively slow- 
growing molds, such as the Aspergilli, from such very rapidly growing forms 
as the Mucoraceae, Trichodcrma, etc. The critical step in this procedure 
is to isolate the slow growing Aspergilli during the first two to three days 
before the whole plate is overrun by the spreading, faster growing forms. 
If the contamination is bacterial in nature, malt extract agar or some other 
medium of more acid reaction should be substituted. Diluting the spores 
in water before streaking is not recommended since this often tends to dis- 
perse the contaminating organism more than it does the desired form; this 
is especially true with bacterial contaminations, actinomycetes, and other 
minute forms. 

Single Spore Cultures 

The isolation of cultures from single spores is a time-honored technique 
with many mycologists, and with many workers it is considered a "must." 
When properly employed, there is much to recommend this practice. There 
are also certain dangers inherent in this procedure, hence we believe it worth 
while to consider the subject at some length and to analyze both its advan- 
tages and its limitations. As generally employed, the primary objective of 
single spore isolations is to secure cultures of unquestionable purity. If the 
operation is skillfully performed and adequately verified, there can be no 
doubt but that the resulting colony will represent a pure culture of a single 
species or strain. The continued purity and physiological stability of such 
cultures, however, cannot be taken for granted. There is no substitute for 
vigilance, and the culture thus isolated must be kept under critical ob- 
servation. 

As a means of purifying a culture, i.e., separating it from foreign forms, 
the single spore method undoubtedly has its place ; but it cannot be recom- 
mended as a means of preserving the morphological and physiological sta- 
bility of a particular strain. One has only to study the reports of Stakman 
and his associates to realize that monospore selection and isolation is not a 
touchstone to strain stability. In fact, they have ably exploited the tech- 
nique to show just the opposite. While we do not have an accumulation of 
data on the Aspergilli and related genera which can compare with that on 
the smuts and other pathogenic fungi, we do have enough information to 
know that one of the best methods of obtaining change in some of the 
Aspergilli is to make a series of successive monosporous isolations. For 
every species and strain there is apparently a normal range of natural 
variation, and by isolating single spores it is often possible to secure certain 



CULTIVATION AND EXAMINATION 43 

progeny which represent essentially the outside limits of such variation. 
This is worth while since it offers one means of securing strains which may 
prove more useful than the parent culture for some particular purpose, 
industrial or otherwise. The single spore method can be of real value in 
purifying a culture. It can be of real value as a means of "dissecting" a 
culture. But once a promising strain has been discovered and its purity is 
established, perpetuation by the transfer of masses of conidia is the best 
safe-guard for preserving, in a constant condition, its morphological and 
physiological characteristics. 

Some of the techniques by which such single spore isolations can be made 
follow : 

(1) Serial Dilation: Spores are thoroughly suspended in water and the 
resulting suspension is subsequently diluted in sterile water blanks in steps 
of 1:5, or 1 :10. One cubic centimeter aliquots from two or three selected 
dilutions, depending upon the density of the original suspension, are added 
to sterile petri dishes. Melted agar at approximately 45° C. is then added 
to these plates, which are rotated to secure uniform mixing of the still liquid 
agar and the diluted spore suspension. The plates are then incubated at 
room temperature and isolates are made from plates showing a limited num- 
ber of colonies which are uniformly separated. Using this technique, one 
cannot be certain that any particular colony results from a single spore, but 
if the original suspension was properly prepared, one can feel sure that more 
then 95 percent of the colonies resulted from single spores. More uniform 
suspensions of spores can be obtained by adding to the suspending medium 
some suitable detergent or aerosol. For this purpose we have successfully 
used sodium lauryl sulfonate in concentrations of 1 : 10,000 or 1 : 100,000 
without apparent harmful effect upon the molds under study. The dilu- 
tion method of securing "single spore" isolations is more rapid than any 
other, and for many purposes it is quite satisfactory. 

(2) Selection and Removal of Individual Spores: Where the investigator 
wishes to be positive that every colony results from a single spore, it is 
necessary to employ some technique combining actual microscopic examina- 
tion with some device for the mechanical removal of selected spores. The 
various types of micro-manipulators are well-suited for this work and, with 
sufficient practice, single spore isolations can be made quite satisfactorily 
with any of these. Dilute spore suspensions in water are mounted on the 
undersurface of a cover slip supported by a glass chamber open at either 
one or both ends. With the aid of mechanical controls and a micro-pipette, 
a single spore is withdrawn from the suspension and ejected upon a suitable 
substratum where the spore develops into a mature colony. 

Single spores can likewise be removed by mechanical cutting devices 
such as those described by LaRue (1920), Keitt (1915), and Lambert (1939). 



44 A MANUAL OF THE ASPERGILLI 

A comparatively thin spore suspension is spread on an agar surface, and 
single well-separated spores are located with the microscope. A small 
cutting device, mounted on a holder screwed into the nose-piece of the same 
microscope, is lowered into the agar and a small block bearing the selected 
spore is removed. This is subsequently transferred to a suitable culture 
surface where the colony develops. 

At the Northern Regional Research Laboratory we have employed a 
somewhat different and simpler method. A thin spore suspension is spread 
evenly over the surface of a firm agar gel that has been specially filtered to 
remove all particulate matter. This is incubated overnight and the spores 
allowed to germinate. On the following day, well-separated sporelings are 




Fig. 11. Single spore isolation. A, A single, well-isolated, germinated conidium 
of Aspergillus niger. B, The same removed on a small agar block and transplanted 
to a fresh agar plate as described in the text, p. 44, X 300. 

located with the aid of a microscope and their positions marked on the 
under-surface of the culture dish. These are then checked with a 8 mm. 
objective and 10 X or 15 X oculars to insure that there are no other un- 
germinated spores in the same area. Using a wide-field binocular of the 
Greenough type and very small micro-scalpels fashioned out of platinum- 
iridium wire (B and S gauge 22 or 24), small agar blocks on which the spores 
spores rest are transplanted to fresh agar plates. Each of these agar blocks 
is then examined again with the 8 mm. objective to insure that the selected 
spore has been transplanted. An experienced worker can isolate from 25 
to 30 spores within a period of an hour. Photographs showing essential 
steps in this technique are presented in figure 1 1 . 



CULTIVATION AND EXAMINATION 45 

Hanging Drop Cultures 

The drop of culture fluid inoculated with conidia and hanging from a slide 
or cover glass into a closed chamber can be incubated and examined readily. 
It furnishes information as to the percentage of conidia which are viable, 
the changes which occur in the germinating spore, such as swelling, bursting 
along definite lines, germination from specialized germ-pores, or branching 
of the germ tube, but descriptions of fruiting structures in such hanging 
drops are worthless in the study of the mold colony or normal fruiting habits 
of the species. 

TEMPERATURE 

The great majority of Aspergilli grow well and sporulate abundantly at 
temperatures of 23° to 26° C. For this reason, most cultures can be incu- 
bated on laboratory tables or shelves, and it is not necessary to give special 
consideration to incubation. There are, however, certain exceptions. 
The large-spored members of the Aspergillus glaucus group, such as A . echin- 
ulatus and A . niveo-glaucus, grow more rapidly and fruit more abundantly 
at 20° than at 24° to 25° C. (Thorn and Raper, 1941). This temperature 
response is especially marked in A. medius: at 18° to 20° C. growth is rapid, 
abundant large conidial heads are produced, and numerous perithecia are 
developed ; at 25° C. and above, growth is restricted, few and smaller conidial 
heads are developed, and only occasionally perithecia are produced (fig. 12). 
On the other hand, the very abundant small-spored members of the A. 
glaucus group, such as A. re-pens, A. chevalieri, and A. amstelodami, grow 
rapidly and fruit abundantly at 30° C. (Thorn and Raper, 1941). In A. 
janus (Raper and Thorn, 1944), two different types of conidial heads are 
produced and the ratio of these types is strongly influenced by temperature 
(fig. 12): at 18° to 20° C. almost all heads are white with clavate vesicles 
and are borne upon long conidiophores ; at 30° C. almost all heads are dark 
green with globose vesicles and are borne upon short conidiophores (see 
species description, p. 187). 

When grown upon suitable media and incubated at 20° C. in the presence 
of light or alternate light and darkness, Aspergillus giganteus produces 
large heads on long conidiophores ranging up to 5 or even 10 cm. In similar 
cultures incubated at 30° C. heads are smaller and conidiophores are uni- 
formly short, rarely exceeding 1 cm. in length, whether the cultures are ex- 
posed to light or incubated in total darkness. A. terreus, A. carneus, and 
A. fischeri thrive at temperatures up to 35° C, while A. fumigatus grows 
well at 45° or even 50° C. In all of these forms growth is more rapid and 
sporulation more abundant at 30° C. than at normal laboratory tempera- 



46 



A MANUAL OF THE ASPERGILLI 




Fig. 12. Influence of temperature upon growth and sporulation in Aspergillus 
medius, NRRL No. 124, and Aspergillus janus, NRRL No. 1787. A , B, and C, Asper- 
gillus medius, 17 days at 12° C, 20° to 22° C, and30° C, respectively ; note maximum 
growth and limited sporulation at 20° to 22° C. D, E, and F, Aspergillus janus, 3 
weeks at 20° C, 24° to 25° C, and 30° C, respectively; note evidence of white heads 
only at 20° C, abundance of white and presence of green heads at 24° to 25° C, and 
green heads only at 30° C. Temperature is the only variable. 



CULTIVATION AND EXAMINATION 47 

tures of 24° to 25° C, although wholly typical cultures are produced at 
both temperature levels. 

In the production of spores for biochemical investigation (see p. 37) and 
in the conduct of such studies themselves, temperature is often very im- 
portant. In work of this kind, it is essential to determine the optimum 
temperature for each organism and process, and then to control this within 
a narrow range in order to secure consistent and reproducible results. 

Temperature is known to be a critical or limiting factor only in a few 
species, but its effect in these is sufficiently pronounced that its influence 
should not be disregarded in any case. 

CULTURE TOOLS AND EQUIPMENT 

Transfer Needles 

In making transfers, we have used with satisfaction for many years a 
Xo. 20 or No. 22 B. and S. gauge nichrome wire thrust into a slender brass 
or stainless steel tube so that only about 15 to 20 mm. are exposed. (The 
tubing employed is of the type used in temperature controls of mechanical 
refrigerators and other thermostatically controlled devices.) The point of 
this wire is then ground down to the sharpness and smoothness of a needle. 
This instrument can be heated to redness in the flame a great many times 
with only the occasional necessity of resharpening. It thus has many ad- 
vantages beside cheapness: it is firmer and takes a better point than plati- 
num; it withstands sterilization in the flame, which promptly destroys the 
usefulness of steel ; and it can be made any size or length to suit the work- 
man's purposes or preferences. 

Loops 

Loops of various dimensions are very useful and can be inserted into 
handles made from small brass or stainless steel tubing as noted above. 
These, too, can be fashioned from nichrome wire, but it has been our 
experience that loops made of platinum-iridium wire possets certain marked 
advantages. While these are not as rigid as nichrome loops, they are much 
more rigid than loops of pure platinum, and they can be re-heated indefi- 
nitely without corroding. Loops of this type will be found especially 
useful in making mass inoculations, such as the seeding of large tubes and 
plates for the production of spores to be used in various types of experi- 
mental work. 

Mounting Fluid 

In the microscopic examination of the Aspergilli it is often satisfactory 
to mount conidia, heads, or other structures in water. It is more generally 



48 A MANUAL OF THE ASPERGILLI 

satisfactory, however, to use some mounting fluid of a composition designed 
neither to swell nor plasmolyze the tissues to be observed. Such a 
mounting fluid was developed by Amann as early as 1896 and has been 
used by mycologists, quite generally, for many years. Its composition 
is as follows: 

Carbolic Acid Crystals (c.p.) 20.0 grams 

Lactic Acid (sp. gr. 1.2) 20.0 grams 

Glycerine (sp. gr. 1.25) 40.0 grams 

Distilled water 20.0 cc. 

The carbolic acid crystals are first liquefied by heating in a water bath. 

The mounting fluid is normally used without the addition of any dye, since 
in the diagnosis of the Aspergilli the natural colors of conidiophores, conidia, 
etc., are very important. If it is considered desirable to stain the tissues 
under observation, it is possible to incorporate into the lactophenol solution 
some coloring substance such as cotton blue, eosin, or some other aniline 
dye. In making mounts of the Aspergilli, it is profitable to wet the material 
first with 70 percent alcohol to drive off air bubbles, and then quickly add a 
small drop of the lactophenol prior to the placement of the cover glass. 

Incubators 

Almost all of the Aspergilli grow well and sporulate abundantly at 
laboratory temperature. There are, however, certain exceptions to this 
general rule (see p. 45), and for this reason it is desirable to have available 
an incubator which can be regulated at temperatures below that of the 
laboratory and others covering different ranges up to 37° C, or even 50° C. 
The type of incubator is not critical and almost any type of cupboard, or 
room, will prove satisfactory if the temperature can be controlled to 
within 1° C. ±, and if the air is neither excessively dry nor humid to the 
point where cotton plugs become moist upon continued exposure. 

Microscopes 

In the study and identification of Aspergilli it is essential to have at one's 
disposal a good-quality compound microscope. If possible, this should be 
provided with apochromatic lenses. In our experience we have found a 3 
mm. objective used in conjunction with either 10 X or 15 X oculars to give 
us magnifications and a degree of definition which is most satisfactory for 
the examination of conidial structures. While it is not absolutely neces- 
sary, it is desirable to have, in addition, a low-power, wide-field dissecting 
microscope covering magnifications from 10 to 40 diameters. In the ab- 
sence of such a microscope a high-quality pocket magnifier provides a good 
substitute. 



CULTIVATION AND EXAMINATION 49 

Photographic Equipment 

A limited amount of photographic equipment is a very valuable aid in 
the study of this or any other group of molds. A camera should be avail- 
able which can be used in conjunction with a compound microscope, and 
to which low-power lenses can be attached directly. In addition to a good- 
quality lens producing pictures above and below natural size, a series of 
Tessar lenses which will provide magnifications of from 5 to 30 diameters 
is extremely useful. With the aid of these, details of colony structure can 
often be recorded which cannot be pictured with the lenses of the compound 
microscope and which are very difficult to describe adequately in words. 
With these low-power lenses it is possible to photograph types of fruiting 
heads, the relative abundance of conidial structures to vegetative mycelium, 
and the relationship between each of the above and sclerotia or perithecia 
when such structures are present. 



Chapter V 
PRESERVATION OF CULTURES 

Any mold that is valuable because it has been used in fundamental 
research work, because it has been found useful in some industrial process, 
or because it is a significant agent in some destructive or pathogenic situa- 
tion should be preserved to insure its identity for subsequent use or refer- 
ence. Identification by description will take the careful worker to the 
group, species, and often to the variety as based upon morphology or some 
conspicuous character, but the reidentification from description of the 
exact organism used in a biochemical investigation or discovered in some 
ecological situation is generally impossible. Culture collections have, 
therefore, developed. The Centraalbureau voor Schimmelcultures (cited 
by them as C.B.S.) at Baarn, Holland, the National Type Culture Collec- 
tion in London, and the American Type Culture Collection at Washington 
are well-known sources of such material. More recently established, but 
containing a greater number of industrially important molds, is the culture 
collection of the Northern Regional Research Laboratory (commonly 
cited as N.R.R.L.) at Peoria, Illinois. The Aspergilli are especially well 
represented, and contained in the collection are almost all of the species 
considered in this manual, together with records which check the identify- 
ing numbers of this collection against the records of the source collections* 
which were brought into it. 

In undertaking a comparative study of the Aspergilli, or any other group 
of molds, it is essential to maintain in a viable state a large number of 
isolates representing diverse species and strains. By this means, it is often 
possible to interpret current isolates and accessions in terms of historic 
types and concepts. In maintaining such a collection of molds, the objec- 
tive should be to preserve viability without growth or germination of 
spores during the storage period. By so doing, the user can reasonably 
expect to maintain his organism without variation, degeneration, or muta- 
tion. A number of techniques can be employed to advantage, the more 
useful of which will be considered in some detail with certain of their 
advantages and limitations noted. 

* Currently contained in this collection are the molds formerly maintained by 
Thorn and Church, and later by Thorn and Raper, in the United States Department 
of Agriculture, Washington, D. C; many of the Mucorineae maintained by Dr. A. F. 
Blakeslee for many years at the Carnegie Institution, Cold Spring Harbor, Long 
Island, New York; a large number of miscellaneous forms from the Harvard Univer- 
sity Collection initiated by Prof. R. Thaxter and more recently maintained by Dr. 
D. H. Linder; and limited numbers of cultures from many other collaborators. 

50 



PRESERVATION OF CULTURES 51 

A gar-Slant Method 

The method most generally employed for maintaining mold cultures, 
and the one which has been successfully used by the writers for many years, 
may be termed the agar-slant method. This involves the periodic transfer 
of spores from old agar slants, or plate cultures, to new agar tubes. The 
composition of the substratum is varied to suit particular requirements and 
groups of organisms. In our work we regularly employ the Czapek's 
solution agar. Few, if any, strains make their maximum growth on this 
medium, but it has been our experience that they maintain exceptionally 
well any characteristic morphological or physiological features which may 
characterize them. It is necessary to know the expected viability of all 
cultures to be maintained, and to gauge the intervals of transfer accordingly. 
With the Aspergilli, transfer every 8 to 9 months is sufficiently frequent for 
all species, with the possible exceptions of A. citrisporns and A. itaconicus, 
and a period of one year is not too long for most forms. In practice it is 
advisable to handle separately the few very short-lived species, and to set 
the regular period of transfer well within the known viability period of the 
remaining forms. Transfer of the general collection at least once each year 
insures a complete survey, within the yearly period, of all strains main- 
tained. New tubes are inoculated at least in duplicate, while triplicate 
preparations afford a desirable margin of safety. The new cultures are 
incubated for 2 to 3 weeks, or until a good crop of spores has developed. 
Incubation at room temperature is suitable for most of the Aspergilli, 
although a few forms such as the large-spored members of the A. glaucus 
group sporulate more abundantly at 20° C. The correctness of the cultures 
is then checked with a wide-field binocular or a 10 X pocket magnifier, the 
plugs are poisoned to preclude any possibility of subsequent contamination, 
and selected tubes are placed in storage. Cultures can be stored for 
reasonable periods at room temperature; certain species will remain viable 
for many years at 24° to 26° C. The viability of most species is materially 
lengthened and the possibility of progressive variation reduced by storage 
at 2° to 4° C. (i.e., above any danger of actual freezing but sufficiently low r 
to prohibit further growth and possible dissociation). Tubes of any de- 
sired size may be employed. We have found lipless tubes 15 by 125 mm. 
to be quite satisfactory since they provide adequate culture surface and at 
the same time require much less storage space than the larger tubes com- 
monly in use. Each culture should be maintained at least in duplicate, 
with the different tubes of each pair stored in separate refrigerators. With 
the accidents and failures of refrigeration, the possible escape of toxic gases, 
or the possible ingress of contaminations that escape the usual inspection, 
the maintenance of not less than two complete series of strains is a necessary 
precaution. If natural conditions, such as temperature and relative hu- 



52 



A MANUAL OF THE ASPERGILLI 



midity, are favorable as at Baarn, Holland, refrigeration may be dispensed 
with, but watchfulness against invasion by mites becomes more important. 
Additional slants should be prepared for cultures which are frequently used. 
Agar slant cultures are convenient for use, easily examined and com- 
pared, and easily replaced (fig. 13 A). They are, however, easily contami- 
nated when handled carelessly. Uneven drying subjects the culture to 




Fig. 13. Methods of maintaining stock cultures as discussed in the text. A, 
Cultures growing on agar slants. B, Cultures preserved in lyophile form; note the 
compact, chalky pellets formed by the dried serum in which the spores are suspended. 
C, Cultures preserved in dry soil. 

extremes of contrast in concentration of media and metabolic end products, 
between the thin edge of the slant and the heavier mass in the bottom of the 
tube. Variations (apparent or real) often appear in the stored cultures 
and these may be propagated in subsequent transfers. As a safeguard, 
stock cultures should be grown in petri dishes after several transfers in tubes, 
thus making possible more complete examination to maintain purity and 
typical morphology. 



PRESERVATION OF CULTURES 53 

Preservation in Lyophile Form 

Studies now in progress at the Northern Regional Research Laboratory 
indicate that many, if not all, of the Aspergilli can be successfully main- 
tained in a dried state for extended periods. Viability tests for a number of 
species including A. terreus, A. niger, A. oryzae, A. flavus, and A. itaconicus 
have been made at 3£ years; while a much greater and wider variety of 
forms has been tested at 20 to 24 months. Positive results have been ob- 
tained with all cultures tested, although comparatively few colonies devel- 
oped from certain strains of A. niger, A. flavus, and the large-spored 
members of the A. glaucus group. Observations are being continued and 
in time information will be obtained as to the feasibility of employing this 
as the principal means of maintaining a collection of molds. It is known 
that many bacteria, especially staphylococci, streptococci, and pneumo- 
cocci can be successfully preserved for periods up to 16 to 18 years (Elser, 
Thomas, and Steffen, 1935; Swift, 1937). Wickerham and Andreasen 
(1942) have presented evidence covering a period of one year which suggests 
the practicability of applying the method to the yeasts. Such information 
as we have to date regarding the molds seems to indicate that the method 
may prove of great significance in two ways: first, as a means of prolonging 
viability, and second, as a means of preserving in viable form spores of a 
particular "generation," or other selected origin, which can be used in 
comparative tests over a period of many months or even years. 

The drying technique employed at the Northern Regional Research Lab- 
oratory is essentially like that described by Wickerham and Andreasen 
(1942) and may be briefly summarized as follows: 

Employing aseptic techniques throughout, the spores from selected 
cultures are suspended in sterile beef, or horse serum. The resulting sus- 
pension is then dispensed into small cotton-stoppered Pyrex glass tubes 
6 mm. by 100 mm. that have been properly labeled with glass-marking ink. 
Approximately 0.05 to 0.1 cc. of the spore suspension is added to each tube 
by means of a long thin-necked pipette. Most of the cotton plug is burned 
away, and the remaining portion pushed down into the tube to prevent 
possible contamination during the drying process. The tubes are inserted 
in rubber sleeves on the manifold, as shown in figures 14 A and B, and low- 
ered into a freezing bath of carbon dioxide ice and methyl cellosolve at a 
temperature of approximately —40° C. The suspension is frozen almost 
instantaneously. The manifold is connected to a vacuum pump and evacu- 
ation and desiccation initiated. Water is removed from the system by the 
insertion of a water-trap immersed in a C0 2 -methyl cellosolve filled Dewar 
flask as shown in figure 14A, or in a column of drierite (anhydrous CaS0 4 ) 
as shown in figure 14B. After a few minutes the temperature of the bath 



54 



A MANUAL OF THE ASPERGILLI 




Fig. 14. Apparatus employed at the Northern Regional Research Laboratory for 
preserving microorganisms in lyophile form. A , Table model of thirty tube capacity, 
utilizing a trap immersed in a Dewar flask filled with CO2 ice and methyl cellosolve 
to collect the water vapor removed from the drying preparations. B, Larger, self- 



PRESERVATION OF CULTURES 55 

surrounding the tubes is raised to approximately —5° C. 1 by supplying 
additional methyl cellosolve that has been precooled to approximately 0° C. 
The bath is maintained at this level continuously until the preparations 
appear thoroughly dry. In drying, the serum contracts slightly to form a 
well-defined chalky pellet. When the pellets are apparently dry, the tubes 
are raised above the bath and evacuation is continued for one-half to three- 
quarters of an hour at room temperature to insure as complete removal of 
water as possible, after which time they are sealed off with a gas-oxygen 
torch (fig. 14 A). On the following day each tube is tested for the presence 
of a good vacuum by means of a high-frequency, spark coil tester, and only 
those tubes which show such a vacuum are retained. Tubes not maintain- 
ing a satisfactory vacuum are very rarely encountered in actual practice. 
Quadruplicate tubes are regularly made for each stock culture, and the 
finished preparations are stored in a refrigerator. 

In recultivating the molds, the tubes are marked with a file scratch, sur- 
face-sterilized, and the tube broken inside a wrapping of sterile cotton. 
The content, which is in the form of a well-formed pellet (fig. 13 B), is then 
dissolved in 1 to 2 cc. of sterile broth or water. This is streaked on agar 
plates and colonies are allowed to develop. New isolations can be made 
within a period of a few days. It is possible, of course, to go directly from 
the lyophile tubes into flasks or other cultures used in actual experiments, 
but generally speaking, much larger quantities of material would need to 
be processed. 

The feasibility of preserving molds in lyophile form over long periods 
has by no means been proved, but results to date are very encouraging. 
Should it be found that spores of molds, like bacterial cells, can be kept 
viable by this method for many years, it will prove ideal as a means of pre- 

1 Wickerham and Andreasen in 1942 governed the temperature at which the sus- 
pension was dried by adjusting the level of the tubes above a bath which was kept 
at a very low temperature. Subsequent to this, Wickerham developed the procedure 
outlined above. 

contained and portable unit of sixty tube capacity (designed by Dr. L. J. Wickerham) 
which utilizes a column of anhydrous calcium sulphate ("drierite") to collect water 
vapor removed from drying preparations. Ai and B\, Glass manifolds; A 2 and B 2 , 
Thermometers; A 3 , Dewar flask containing water vapor trap immersed in CO2 
ice and methyl cellosolve; B u Column of drierite; A^ and B b , Freezing bath con- 
taining CO2 ice and methyl cellosolve in which preparations are immersed for 
quick freezing and subsequent temperature control; Ae, and Be, Vacuum pump; At, 
Vacuum guage (mounted at opposite end of apparatus shown in figure B) ; As and B s , 
Gas-oxygen torch for sealing off dried preparations; B 9 , Terminals on which finished 
tubes are mounted to be tested for presence of good vacuum by means of a high fre- 
quency spark coil tester (not shown); Bio, Oxygen tank; B u , Screw lift for raising 
and lowering manifold and attached tubes (In figure A , manifold is raised and lowered 
manually and locked into position by means of a wing bolt). 



50 A MANUAL OF THE ASPERGILLI 

serving large culture collections. It possesses certain marked advan- 
tages : 

(1) There is no possibility of contaminants entering the sealed prepara- 
tions. 

(2) The investigator recultivating the molds starts with the actual spores 
contained in the original suspension. 

(3) The space required for storage of a large number of lyophile prepa- 
rations is much less than for any other type of culture (fig. 13 B). 

Preservation in Soil 

Soil has been successfully employed as a means of preserving vigorous 
stock cultures over long periods. As early as 1918, Barthel (Cent. Bakt. 
II, 48: 340-49. 1918) reported the successful maintenance of yeasts and 
bacteria in this medium and modifications of this technique are now em- 
ployed in many laboratories for preserving bacteria. Greene and Fred in 
1934 compared cultures of various molds preserved for two years in soil 
with the same strains continuously maintained on malt extract and malt 
extract-potato-glucose agars and on bread. In their experience, soil prepa- 
rations were most satisfactory, and Professor Elizabeth McCoy (personal 
communication) has recently reported cultures of A. sydowi preserved in 
this manner to be viable after nine years. Since the publication of Greene 
and Fred's work, the soil method has been rather generally used by the 
Wisconsin group as a means of preserving valuable stock strains of molds. 
Furthermore, it is known to have been successfully employed during the 
past two years by a number of laboratories to maintain cultures of penicillin- 
producing molds in a high and uniform state of productivity. The soil 
substrate used by Greene and Fred was prepared as follows: 

"To air-dried orchard loam soil (Miami silt loam) sufficient water is added to 
bring it to a moisture content of about 20 percent. The soil is then transferred in 
convenient amounts (about 5 grams on a dry basis) to ordinary half-inch (1.27 cm.) 
culture tubes. The tubes are plugged with cotton and given four 3-hour steriliza- 
tions at 15 pounds per square inch (1 kg. per sq. cm.) pressure on alternate days, and 
tested for sterility by addition of yeast-water-glucose broth to tubes selected at 
random. The tubes are then inoculated with 1 cc. of a heavy spore or mycelium 
suspension of the desired mold and kept at room temperature. That there is appre- 
ciable growth and sporulation on the soil can usually be ascertained without difficulty 
by direct microscopic observation. While the addition of nutrient to the soil may 
bring about somewhat greater growth, it does not seem to enhance the keeping quali- 
ties of the cultures. 

"It has been found possible to preserve on soil mold stocks used for large-scale 
growth— namely, Aspergillus fischeri, A. sijdowi, and Penicillium chrysogenum— for 
over 2 years without loss of their essential and desirable characters. . . . Moreover, 
the gross colony characters have remained much more constant than did those of the 
corresponding cultures maintained in the usual way on agar slants. The soil cultures 



PRESERVATION OF CULTURES 57 

can be recovered as required simply by streaking some of the soil particles on fresh 
agar slants. 

"It is not in all cases advisable to depend on soil alone for the preservation of 
valuable stocks, but reserve stocks may without difficulty be prepared on soil, and 
the writers believe that in many instances soil will be found to be an excellent medium 
for maintenance, with a minimum of change over long periods of time." 

During the past two years the soil method has been used at the Northern 
Regional Research Laboratory with but minor modifications of the tech- 
nique cited above (fig. 13 C). Its principal advantages lie in the fact that 
(1) the viability of strains is apparently lengthened, and (2) from a single 
stock tube, opened with proper care, repeated cultures can be started 
simply by removing some of the soil particles to suitable substrata. 

Vegetable Substrata 

The oriental fermentation industries maintained their inoculating mate- 
rial as selected rice or soybeans upon which the mold had been grown under 
favorable conditions to produce maximum quantities of spores. This nu- 
trient, dried and packaged, was stored and sold under the Japanese name 
"Koji". Samples examined after several years showed excellent viability. 

Bainier was a pharmacist. He distributed licorice root in sections 5 to 10 
mm. in diameter and 5 to 8 cm. in length in test tubes, sterilized them, and 
kept his cultures regularly for years upon them. Tested by us, the method 
was a very satisfactory laboratory practice. American mycologists have 
successfully used bean stems for the purpose. Apparently any organic 
material which provides frameworks of cellulose enmeshing sufficient nutri- 
ents to support mold growth without complete breakdown of the mass may 
be used. 

CONTAMINATION 

Mixed Strains 

In the routine conduct of cultural work, contaminations of cultures of one 
species of Aspergillus by other species, or species of other genera, is very 
common. The conidia of most molds are exceedingly light and are carried 
freely in the air. Entire exclusion of such contamination is difficult. In 
dealing with contaminations, several problems arise and different proce- 
dures are possible. A colony of a single Aspergillus, well established, 
usually inhibits the growth of other species developing in the immediate 
vicinity. Even if invasion occurs, the effects are commonly so distinct as 
to leave little doubt as to the limits of the different forms. When, however, 
the contamination with spores or mycelium is carried in the inoculum and 
so placed as to germinate in intimate contact with the organism desired, (1) 
the species may sector out, and hence be easily recognized, or (2) the colony 



58 A MANUAL OF THE ASPERGILLI 

resulting may assume the character of either organism with the other 

present only as an inconspicuous, even unrecognizable, mycelium with 

dwarfed heads, yet continue present for a long period. In this way the 

retarded species may suddenly reappear in some later transfer upon media 

favorable to it. In other cases, the dominant species may grow and fruit 

in an erratic manner that is deceptive in suggesting a reaction to the medium 

r other physical factors. Again, in less common cases, the two may grow 

jid fruit together without apparent inhibiting effects. This is the most 

lifficult form of mixture. 

Mixtures are sometimes encountered in which mycelia, sterile under all 
conditions tested, become so intimately mixed with the mycelium of an 
Aspergillus or Penicillium as to persist through many generations without 
apparent effect in the earlier states of growth of the Aspergillus, but develop 
as overgrowths of sterile hyphae in very old cultures. Many of these forms 
can be isolated, but they defy identification because of an absence of diag- 
nostic characters. Such sterile mycelia may arise from the species studied, 
but unless such origin can be definitely proved, they must be regarded as 
contaminants. Great care is necessary in interpreting cultures producing 
sterile overgrowths, since the contaminating organism, or non-sporulating 
variant of the same strain, may induce marked changes in the physiological 
activity of the species supposedly pure. 

Secondary Growth 

In many species, part or all of the conidia produced by a colony germinate 
and cover the primary mycelium with more or less abortive hyphal growths, 
many of them unrecognizable unless traced by their origin. Again spores 
floating on the surface of a globule of transpired fluid may germinate, their 
hyphae interlace and a hollow ball surrounded by felted mycelium produce 
structures which probably account for reported perithecia without asco- 
spores. Such overgrowths, being irregularly produced, interfere with one's 
judgment as to the whole character of the colony. 

Replacement by Other Species 

Other species of fungi invade mold cultures and some of them become so 
intimately associated with the mycelium and conidia of particular species 
that it is difficult to eliminate them by the ordinary method of transferring 
spores with a loop or wire to streaks or stabs. Proper dilution culture 
presents the possibility of elimination but demands careful examination 
and selection from the resulting colonies. The progressive replacement of 
a particular species, by invading organisms, is constantly encountered in 
examining cultures passed from laboratory to laboratory. The original 
organism may disappear without detection if the displacing species bears 



PRESERVATION OF CULTURES 59 

a superficial resemblance to it, or if no detailed examination of successive 
transfers is made by a worker really familiar with the specific characters of 
the stock strain. The physiological or biochemical investigator, obtaining 
such a culture assumed as correctly named, will be seriously misled in the 
interpretation of his experimental results. No culture should be used for 
such an investigation without being fully identified at the outset and having 
its proper appearance and reactions sufficiently studied to insure the relia- 
bility of the results during the progress of the work. In other words, before 
undertaking to do biochemical or physiological work, sound scholarship 
calls for adequate precautions in the study and identification of the original 
material supplemented by such mastery of its morphology and variability 
as will insure its maintenance in proper condition. 

Mold Disease of A . niger 

A mold disease of Aspergillus niger is common in cultural study. The 
colonies of the Aspergillus are overrun with an olive-green Penicillium 
belonging in the Biverticillium group close to Penicillium rugulosum Thom. 
This mold invades the mycelial felt, winds its hyphae within and about the 
conidiophores, and fruits in a radiating series of short-stalked penicilli 
surrounding the heads and upper halves of the conidiophores. This is 
beautifully illustrated in figure 15, made by Edward Yuill and sent to us by 
his brother John L. Yuill of Yorkshire, England. The black fruiting surface 
may be completely covered with the olive-green conidial masses of the 
Penicillium. If A. niger is grown in trays for acid formation, spots infected 
in this way may be killed and disintegrated, thus interfering with fermenta- 
tion, while the infected areas may be seen to drop out when the blanket or 
felt of A. niger is lifted from the surface of the liquid. Other species when 
inoculated have been irregularly affected, some strains of the same species 
were attacked, whereas others were apparently immune. 

Bacteria 

Freedom from bacteria is essential to uniformity in the appearance and 
in the reactions of molds. Colonies infected with bacteria may be un- 
changed in character, but usually show marked physiological differences 
when compared with colonies free from contamination. Associative action 
may have important effects upon both organisms. Sartory (1920) dis- 
cussed without identifying a species producing ascospores, but only when 
accompanied by a particular bacterial associate. Nevertheless, a symbiotic 
colony must not be allowed to masquerade as a pure culture representative 
of a species. The next contaminated colony of the same species of mold 
but with different bacteria may present a very different picture. These 
conditions have been met often enough to make the emphasis upon freedom 
from bacteria essential in study of this group. 



60 



A MANUAL OF THE ASPERGILLI 



Mites 

Mites are very common in rotting vegetables, especially in partly dried 
condition, in dried meat products, in hard cheeses, and in organic soil 
masses. They are thus common associates of molds as they occur in nature, 




Fig. 15. Penicillium sp. parasitic on Aspergillus niger, X 165. (Photograph by 

Edward Yuill) 



hence the worker who handles moldy substrata, in isolating his organisms 
must constantly watch for them. In size, mites are commonly just about at 
the limit of visibility by the unaided eye. One accustomed to them will 
detect them readily; but until seen and the appearance of their depredations 



PRESERVATION OF CULTURES 61 

understood, they can pass unnoticed for considerable periods by persons 
who are otherwise good culture workers. Mites will crawl from petri dish 
to petri dish, leaving behind them a trail of bacterial and mold contamina- 
tions, as well as streams of eggs which develop rapidly into more mites that 
actually destroy the colonies. Since mites have preferences as to food, 
some species are invaded and others avoided. To reach an attractive food 
supply a mite will frequently go through a cotton plug as ordinarily made 
and occasionally seems to get through even a paraffined plug. As a factor 
in the mixing of strains of molds in a laboratory collection, mites must not 
be ignored. 

Similar mixing and contamination occurs frequently whenever a labora- 
tory becomes infested with ants, roaches, or other insects. 

Poisoning Cotton Plugs 

Mites: Entire elimination of mites by sanitary measures is possible but 
often not attained. As a precaution in the preservation of stock cultures, 
some scheme of poisoning should be used. One of these formulas consists 
of dipping the tips of the cotton plugs in a solution of the following com- 
position: 

95% alcohol 95 cc. 

Bichloride of mercury 0.5 gm. 

Glycerine 5 cc . 

Color with any aniline dye. 

Care must be taken that the solution does not come in contact with the 
colony. The cultures must be allowed to develop into typical colonies 
before poisoning. An antiseptic formula for the purpose needs alcohol to 
insure penetration of the plug, a poison to destroy the mites, glycerine to 
prevent the crystallization of the poison as the alcohol evaporates, and the 
dye to insure the destruction of the cotton plugs when removed from the 
tubes. In our own experience we have consistently made it a practice to 
wipe off the outside of all culture tubes with the above, or some other 
sterilizing solution, and to poison all plugs before cultures are replaced in or 
added to the collection of stock cultures. 

Molds: Under humid laboratory conditions, cotton plugs, especially if 
made from the absorbent type of cotton, absorb moisture. Careless han- 
dling in preparation and care of such plugs often adds enough nutrients to 
support growth. Steam sterilization tends to distribute nutrients. Dirt, 
bacteria, and molds fall from the air upon the exposed portion of the plug. 
Handling detaches spores from the colony within so that both ends of the 
plug are commonly well seeded. Spore germination, therefore, may begin 
at either or both ends. Outside molds may grow through and drop into the 



62 A MANUAL OF THE ASPERGILLI 

culture, or the culture itself may grow out through the plug and contaminate 
other cultures or experiments. Surface sterilization of the outside of the 
tubes and poisoning the plugs takes care of molds, as well as mites. 

Spraying: Oily sprays, as selected fractions from petroleum, avail- 
able from commercial sources, even kerosene, distributed with a "gun" 
that produces a mist penetrating and filling all cracks, crevices, open spaces 
among apparatus or furniture and clouding the whole atmosphere of the 
laboratory, have been found effective in carrying down mold spores and 
bacteria from the air and ridding the laboratory of mites, insects, and 
vermin. 

PRESERVATION OF DRIED SPECIMENS 

Mold cultures lose many of their characteristic and diagnostic features 
upon being dried. Nevertheless, dried herbarium specimens serve a useful 
purpose in preserving type material which might otherwise be lost. Details 
of morphology are often difficult to establish from such material, but group 
characteristics are preserved and over-all colony appearances can be recog- 
nized after many years. The retention of culture tubes or petri dishes 
containing such dried specimens constitutes a reasonably satisfactory means 
of preservation, and the material contained therein approximates as nearly 
as is possible the cultural picture of the growing colony. Glass tubes and 
dishes, however, are cumbersome and easily broken, hence may prove 
unsatisfactory if frequent handling is necessary. For many years we have 
employed an alternate technique with generally satisfactory results. Rep- 
resentative portions of colonies grown in petri dishes are cut with a large 
cork borer, lifted out with a spatula, and dropped into paper pill boxes 
where they are allowed to dry. These can then be stored in larger boxes 
or attached to herbarium sheets for filing. The boxes should be provided 
with tight-fitting lids, and for greatest convenience should measure approxi- 
mately l\ inch in diameter. Aspergilli stored in this way prove useful in 
many comparative studies. They cannot, however, under any condition, 
take the place of carefully handled living cultures. 



Chapter VI 
VARIATION 

The Aspergilli are a variable and mutable group of fungi. They are 
characterized by great diversity and variability as they are isolated from 
nature, and an increasing amount of evidence shows that they can be made 
to vary, or mutate, in the laboratory by subjecting them to a number of 
different imposed stimuli. Frequently the same types of mutants or vari- 
ants ultimately result under both natural and artificial conditions. Never- 
theless, it is believed desirable to consider somewhat separately variations 
and mutations resulting from natural causes and those resulting from 
imposed stimuli. 

Definition of Terms 

Before entering upon a discussion of variation, either natural or induced, 
it is important to define certain terminology which is to be employed. It 
is recognized that our definitions will not agree in all cases with those of 
earlier workers, nor do we expect that all subsequent investigators will 
accept those which we propose. If the meaning in the present discussion 
is clear, our purpose will have been served. 

(1) The term mutant, or mutation, is used to designate a strain whose 
source is actually known and can be verified. Furthermore, it is limited to 
those substrains which originated as sharp breaks from parent cultures 
(usually interpreted as gene mutations), and in successive culture genera- 
tions retain their distinguishing characteristics unaltered. This may or 
may not have a taxonomic connotation. Upon occasion it is used in essen- 
tially the same sense as "variety". To illustrate, A. nidulans mut. albus, 
A. fumigatus mut. helvola, A. niger mut. cinnamomeus, etc., are used as 
Latin names to designate forms which differ from the parent species in 
certain striking details. In other instances it is used to identify a type of 
change, rather than to designate a particular and isolated strain resulting 
from such change. It is considered correct to refer to artificially produced 
albino, yellow, and buff-colored strains of A. terreus as mutations (see p. 
75), since they are constant in character and are known to have originated 
from a cinnamon-colored parent culture, wholly representative of the 
species; and we believe it represents good judgment to refrain from assign- 
ing Latin designations to each of them. The term mutation, then, refers 
to altered strains of constant character and known lineage, whether or not 
the y are given Latin designations. 

63 



64 A MANUAL OF THE ASPERGILLI 

(2) The term variant, or variation, is used loosely and reference to it in 
this manual is not, in all cases, entirely consistent. In general, however, it 
is applied to subcultures, or strains, arising through gradual change from 
well-defined strains of identifiable species. The characters of a variant, 
then, are not generally stable but subject to continued change and further 
variation. As used by us, the term has no taxonomic implication and can 
be considered essentially synonymous with the term "saltant" which is so 
commonly employed in reports on variation in the Fungi Imperfecti. 
Variants frequently appear as colony sectors, overgrowths, or other local- 
ized areas of changed appearance or texture. When isolated in pure cul- 
ture, they may or may not retain their distinguishing characteristics. 

NATURAL VARIATION 

Cosmopolitan species and groups of Aspergilli show adaptability to wide 
ranges of environmental conditions. As these molds are isolated from 
nature, variation among the members of any species, series, or group is 
regularly encountered. Such variations commonly differ in degree rather 
than in basic characters, and one can distinguish a series of intergrading or 
bridging forms. Even striking isolates are often unmistakably allied with 
some well-defined species or group in this manner. Such different but 
intergrading forms arising in nature can be considered as natural variants. 
Natural variants of a similar kind can frequently be obtained in laboratory 
culture by selective isolation and cultivation from sectors or other areas of 
atypical growth, or by single spore isolations. Distinction must be drawn 
between differences in appearance, morphology, and habit of growth result- 
ing from inherent differences between strains, and alteration in colony 
character in response to changes in the composition of the culture medium 
or other environmental factors. Rigid comparative culture is often neces- 
sary to distinguish between the two. For the present discussion, we are 
concerned with differences that are more fundamental than direct tempo- 
rary responses to artificial stimuli (ecads); but the latter, unless carefully 
evaluated, may appear no less real. Rightly or wrongly, Blochwitz (1930, 
p. 247) comments that .4. flavus, 1 in specimens collected in the Botanical 
Garden at Buitenzorg, was called A. penicillopsis (Henn.) Rac; in the 
Botanical Garden at Singapore, *S. vitellina Ridley; in India, S. corolligena 
Massee; and in Columbia, .4. delacroixii Saccardo. 

Occasionally isolations are made from laboratory cultures which represent 
sharp "breaks" from the parent strain, and since they are constant in subse- 
quent culture, they may be considered as true mutations. Representatives 

1 The name of the original describer is only used for specimens or strains in culture 
which were definitely attributed to the describer. A. flavus, A. niger, A. terreus, etc., 
are series concepts as used here. 



VARIATION 65 

of such natural mutations which originated in the absence of any artificially 
imposed stimuli are YuilPs A. fumigatus var. helvola (1939), A. nidulans 
var. albus (1939), and Cladosarum olivaceum (1938). 

Intra-strain Variation 

A certain amount of variation can be expected to occur in any given strain 
of Aspergillus. In certain species and strains this is very limited and cul- 
tures can be re-cultivated repeatedly upon a variety of media, and at 
different temperatures and H-ion concentrations without evidences of visi- 
ble change other than those resulting from the immediate effects of the 
altered environment. Many strains of Aspergillus niger are characterized 
by such comparative stability. Other species and strains are subject to 
continual variation with differences in character and rate of growth appear- 
ing rather abruptly as sectors or overgrowths, or gradually developing as a 
progressive alteration in the general aspect of the whole culture. By suc- 
cessive and selective subculturing, strains of A. alliaceus and A. ochraceus 
showing a marked difference in sclerotium production can be obtained. In 
like manner, strains of A. itaconicus Kinoshita can be secured which are 
almost completely sterile upon all media tested. The same is true of A. 
granulosus Raper and Thorn, A. flavipes, etc. 

Working with a strain of the ascosporic species, A.fischeri, Greene (1933) 
isolated 448 single spore cultures and among these found variant progeny 
of two main types: (1) cultures producing very large, scattered perithecia 
as opposed to the typical picture of many small perithecia, and (2) cultures 
producing conidial structures in profusion, but forming few perithecia and 
these tardily. The second type was fairly stable, whether derived from 
single ascospores or conidia. The first type was variable, in some cases 
reproducing the characters of the variant parent, in others reverting to the 
character of the original stock culture. 

Hansen (1938) and Hansen and Smith (1932) have studied many of the 
Fungi Imperfect! rather exhaustively and report that single strains of these 
fungi are basically composed of a mycelial (M) type and a conidial (C) type. 
By proper techniques the two forms can be separated and recombined at 
will. While it has not been explored as yet, the possibility exists that the 
same condition may prevail to a limited degree in the Aspergilli, and that 
this may account for a certain amount of the intra-strain variation en- 
countered. 

Back of variability in molds, many lines of discussion have been devel- 
oped. Buller (1933) has frequently called attention to the unmeasured 
possibilities of nuclear and cytoplasmic disturbance from the commonly 
observed phenomenon of anastomosis. Vegetative hyphae belonging to the 
same mycelium (mycelium derived from a single spore), or different mycelia, 



66 A MANUAL OF THE ASPERGILLI 

throw out branches which fuse without showing any other sign which might 
suggest a sexual process either before or after fusion. Anastomosis is 
usually observable, if at all, in the rapidly growing area where many spores 
placed as an inoculum are developing into one colony. By transferring 
large numbers of spores from an old culture to a new one, most of the 
Aspergilli studied by us have shown fairly consistent repetition of colony 
characters and conidial morphology and have been maintained for a long 
time with little or no observable change. Other organisms handled in the 
same manner have not been successfully maintained with the morphology 
originally studied. Differences in behavior can be attributed to variability 
between strains. 

Intra-species Variation 

Variation within the species is very prevalent and is well marked in many 
cases. When a large number of isolates of any particular species or series 
is collected, one can regularly expect to find among them wide variations in 
color, amount of sporulation, and in their general habit of growth. Usually, 
however, such variation is graduated, and strains representing various 
intermediate steps between the extremes are to be expected. While it is 
by no means unique, we may use A. terreus as an example, since a very large 
number of strains belonging to this species have been isolated and observed 
in plate and tube cultures during the past two years (fig. 16). Colonies of 
the type strain, and of the great majority of isolations made from nature, 
are plane, cinnamon in color, very heavy sporing, with conidial heads arising 
directly from the substratum in an even and close stand. An occasional 
isolate is much brighter in color, approximating xanthine orange (Ridgway, 
PL III), but in all other respects it is fairly typical. It is believed to repre- 
sent a form such as that described by Blochwitz (1934) as A. boedijni, and 
in the present manual we have designated it as A . terreus var. boedijni. In 
each of the above cases, the production of abundant fruiting structures 
follows closely the advancing margin of the growing colony and there is little 
or no continued growth of mycelium except in the marginal area. Certain 
other strains are quite floccose; conidial heads are typical in form and color 
but are greatly reduced in number and are borne upon aerial hyphae. Shih 
(1936) was probably working with such a culture when he described the 
variety A. terreus var. floccosus. We feel that the forms are sufficiently 
distinct to warrant maintaining his variety. Still other strains possess 
abundant but fairly close-, rather than loose-textured mycelia and bear 
abundant but very pale buff-colored conidial heads. The vegetative my- 
celium in this form is bright yellow and for this reason we have designated it 
as A . terreus var. aureus n. var. (see p. 198) . If one should examine only the 
type strain and these three atypical forms, it is entirely probable that one 



VARIATION 



67 



would describe them as four distinct species. Actually, however, they are 
not sufficiently distinct to warrant specific rank, for they represent only 
extremes of variations along three divergent lines with numerous inter- 




Fig. 16. Intraspecies variation in Aspergillus terreus. A, A. terreus, typical 
strain NRRL No. 265, characterized by heavy conidium production and colonies 
cinnamon in color. B, A. terrreus var. boedijni, NRRL No. 680, characterized by 
deeper colonies and conidial heads near xanthine yellow. C .A . terreus var. floccosus, 
NRRL No. 1921, characterized by loose floccose colonies and spore heads light 
pinkish-cinnamon in color. D, A. terreus var. aureus, NRRL No. 1923, character- 
ized by yellow floccose colonies and comparatively few cream to buff-colored conidial 
heads. 



grading strains aligning them, almost without interruption, with the typical 
form itself. 

Another series of variants in our collection shows gradation from the 
usual radiate head and conidiophore of A . sydowi to the simple mono-verti- 
cillate penicilli of Penicillium restrictum Abbott. The tendency to form 



68 A MANUAL OF THE ASPERGILLI 

reduced conidial apparatus is observed in all of the strains of A. sydowi 
examined. Over a period of many years variants have exhibited all 
gradations in colony appearance from typical A. sydowi to the aspect of P. 
restrictum of Abbott except for the presence of an occasional conidiophore 
and head of .4. sydowi. Repeated cultural tests exclude contamination. 
These variants occur in nature, they produce abundant conidia, and they 
undoubtedly maintain themselves successfully in the field. 

Aspergillus fumigatus presents a somewhat similar condition. In this 
species, typical cultures produce heavily sporing, velvety colonies that are 
dark green in color and show almost no aerial mycelium. Other strains 
commonly isolated from nature produce very floccose colonies and bear 
comparatively few conidial heads (fig. 37). These heads, however, are 
typical in form and in dimension. Strains possessing this contrasting 
character are relatively stable in culture, but in this species, as in A. terreus, 
all degrees of intergradation are found between this floccose type and strains 
entirely typical of the species. The same story is repeated in other species. 

Appreciable variation can normally be expected among the isolates of 
any of the very abundant and cosmopolitan species. Such variant strains, 
however, are the exception rather than the rule, since the great majority of 
isolates are quite typical of the species. Attention was called to this fact 
in our study of the A. glaucus group (1941). 

When grown in comparative culture, strains successively isolated as repre- 
senting a particular species usually show enough difference to give each 
strain a kind of individuality. Exact identity, point by point, is not ex- 
pected. Such strain variation may be incidental and unimportant, or it 
may be correlated with activities which make one strain a valuable agent 
in an industrial process and the other worthless. 

Intra-group Variation 

Inside the different groups of Aspergilli one normally finds somewhat 
similar but wider variations than those seen among the strains constituting 
any particular species. To what degree species in nature have developed 
by mutations and by progressive variation can only be guessed. We do 
know, however, that the species within a group, like the strains or varieties 
within the species, are regularly bridged by intermediate forms which render 
it difficult to establish sharp and immutable lines of separation. One can 
almost cite it as a rule that the definiteness with which one regards a species 
is inversely proportional to the number of strains of that species which have 
been examined. Still species are necessary as guide-posts — as fixed points 
around which closely related organisms showing a certain but limited 
amount of variation can be grouped. 

The Aspergillus flavus-oryzae group can be taken as illustrative of the 
type of variation to be expected within a group of the Aspergilli. Thorn's 



VARIATION 69 

culture No. 113 (XRRL No. 447) of A. oryzae, received from Baarn and 
believed to stem from Cohn's original strain, is a very floccose, loose-textured 
culture bearing comparatively few, small, light yellow to tan heads. Co- 
nidiophores are long, ranging up to 2 to 5 mm., and are very thin- walled. 
There is only a trace of green even in individual heads, and in general aspect, 
the culture normally shows no green color. Aspergillus flavus in its typical 
form is not floccose and is very heavy sporing. Conidiophores arise directly 
from the substratum in a close stand, are usually 1 mm. or less in length, 
and are comparatively heavy walled. Colonies are regularly in yellow- 
green shades and range from light to comparatively dark green (see species 
description). A. parasiticus Speare goes even farther. Colonies are very 
dark green in color. Conidiophores usually range from 200 to 400m in 
length, and sterigmata are typically in a single series, whereas they may be 
in a single or double series in A . oryzae and A . flavus. In the opposite direc- 
tion, but markedly different from A. oryzae is A. effusus Tiraboschi. Typi- 
cally this is very floccose and comparatively light sporing, with heads borne 
upon short conidiophores which arise from the loose aerial mycelium rather 
than from submerged mycelium in the substratum. 

The cultural pictures of these species are fairly characteristic. Yet, it is 
practically impossible to take a large collection of 100 or more strains and 
separate them into these species with any degree of confidence or satisfac- 
tion. The difference between A. oryzae and A. flavus is bridged completely 
by a series of intermediate forms showing all degrees of variation between 
the two strains selected as typical. Xomenclature in this group is then 
further complicated by the fact that among the great collections of these 
forms obtained from the Orient, and designated A. oryzae, the majority of 
forms are somewhat intermediate between A. oryzae and A. flavus as de- 
picted above. A similar series of intermediate forms bridges completely 
the gap between A. flavus and A. parasiticus. There is no sharp line of 
demarcation between any of these species, still they are not one and the 
same, and to attempt to lump these diverse forms together into one species, 
as Xeill (1939) has done for the ^4. glaucus group, intensifies rather than 
reduces the difficulties encountered. 

Intra-group variation is also particularly marked in the .4. niger group. 
During an extended period of study and observation of molds in culture, 
Biourge, who was a discriminating collector, accumulated 63 strains of 
black Aspergilli which suggested sufficient individuality to be deemed 
worthy of further study. These were turned over to Mosseray when he 
entered Biourge 's laboratory. He assumed that he had before him all of 
the black Aspergilli possible to collect and, knowing that Biourge had 
selected each of them because it seemed to have some special character, he 
undertook a taxonomic study to define those characters and to organize 
them into a systematic presentation (1934a). His paper lists 35 species of 



70 A MANUAL OF THE ASPERGILLI 

which 25 were either described as new species or new combinations. His 
findings in A. niger are fairly illustrative of the same type of study in other 
groups (compare Thorn and Currie [1916] for A. niger; Thorn and Church 
[1921] for A. flams. 

Mosseray based his primary separations upon conidial sizes, shapes, and 
markings, while secondary and tertiary separations were based upon co- 
nidiophore lengths and the characters of colonies in tube cultures on 
Biourge's "Raulin-neutre gelose" — a variation of the classic Raulin solution. 
Biourge and Simonart demonstrated the entire series to one of us (C. T.) 
showing how wrinkling and granulation of mycelium, intensity and changes 
of secreted color, shades of color in the conidial area, lengths and propor- 
tions of conidiophores and heads, and their distribution over the mycelium 
gave to each strain an individuality which had been repeated in successive 
cultures over a considerable time. We raised just one question, "What 
would you do with the next thousand?" 

The large majority of all isolations of black Aspergilli conform within a 
range of minor variation with the general van Tieghem concept, — i.e., 
black-brown colonies with conidiophores and heads giving the general 
structure and measurement of parts found in the classical description. 
Then, in contrast with these, there are shades of colony color from the coal 
black of A . carbonarius through shades of purplish-black to the brown of A . 
ferrugineus Fuckel or to the lighter shades of Schiemann's mutants (pp. 
223-224) . Some of them produce no colors in the substratum and reverse 
of the colony; some show yellow in traces; others are persistently deep 
orange, giving the whole a yellowish appearance. Or, again, the agar and 
mycelium may develop a red-brown or "mauve" shade of violet. 

Conidiophores in the usual type of culture reach nearly enough the same 
length to give the effect of a field of grain. But their length may be quite 
short and the heads seem to be borne directly on the substratum, or they may 
be several millimeters in length with the heads borne well above the sub- 
stratum and correspondingly large. Between these extremes, every varia- 
tion can be found. Conidiophores may be scattered thinly over the 
vegetative mycelium, collected in a zone at the border, or crowded in 
the center. 

The vegetative mycelium may grow as a flat felt (plane) or may be vari- 
ously wrinkled, sulcate, or buckled. In a smooth or plane colony the 
mycelial cells seem to stop growing early — the colony extends only at the 
margin. In the plane colony intercalary growth (i.e., the formation of new 
cells in the filament, or the lengthening of the old cells), and the production 
of new branching ceases. Such mycelia ordinarily produce one crop of 
conidial heads, beginning at the center of the colony and progressively 
developing toward the margin until the medium is exhausted or some 
inhibiting factor paralyzes growth. Marginal growth in such a colony 



VARIATION 71 

often shows longer and fewer conidiophores and larger heads than in the 
central area. 

Within the group with the usual structures still recognizable, many 
variants with contrasting features appear. Heads with very long primary 
sterigmata, which are sometimes septate, appear in A. carbonarius (Bainier) 
Thom, A. pulchella Speggazini, or A. tubingensis Mosseray. The primary 
sterigmata may grow out into sterile filaments as in Mosseray 's figure for 
A.ficuum (Reich.) Henn.; much more commonly, some primary sterigmata 
grow out as tiny conidiophores and produce little heads, often consisting 
only of a cluster of simple sterigmata and conidial chains. Thus, sterig- 
matic changes may run from the simple sterigmata of A. japonicus Saito, 
A. luchuensis Inui, or A. malvaceus Mosseray, where only some are double, 
to other species showing the widest range in length and arrangement. 

Another group of variants show marked suppression of the ordinary 
structures expected. Strains in which conidiophore formation has been 
reduced or almost suppressed have been studied in continuous culture. 
Such colonies showed an occasional long conidiophore and large head, con- 
forming to the A. niger pattern, produced at the end of the colony growth 
period. Meanwhile the mycelium was fully covered with irregularly 
branching hyphal elements bearing single sterigmata variously placed, 
groups of sterigmata, or penicilloid clusters of sterigmata each bearing a 
short chain of conidia showing the characteristic markings of the group. 
Transfers from the simplest form developed the complex or A. niger ele- 
ments. Transfers from the large heads brought a recurrence of the re- 
duced type of fruiting. No method of selection tried brought back the 
typical A. niger aspect, and cultures of this type appear to represent de- 
generate forms. 

It would appear, then, that a general type or morphological picture when 
found dominant in large numbers of natural isolates can be regarded as 
typical for a species of Aspergillus. It is recognized that marked diver- 
gences from such types occur under the unrecorded stimuli of nature. 
Some of these forms succeed in establishing themselves as permanent 
elements of the microflora and thus become successful as species, or va- 
rieties. Others do not digress quite so markedly and thus constitute 
intermediate or bridging forms. At the same time marked changes can 
be induced by the application of artificial stimuli. Where the origin of 
such altered strains is known, they are commonly regarded as mutants. 
Were they isolated directly from nature and their previous history not 
known, it is probable that they would be considered as separate varieties 
or even species. 

Natural Mutation 

While most of the mutants which we recognize as such have originated 
in the laboratory as the result of certain artificially imposed stimuli (or 



72 



A MANUAL OF THE ASPERGILLI 



drastically altered conditions of growth), and hence can properly be termed 
induced mutations, a number of well authenticated cases of natural muta- 
tion are known. In 1939 Edward Yuill described as .4. fumigatus var. 
helvola a buff-colored mutant of this species isolated by him in 1937 (fig. 




Fig. 17. Natural mutations. A, Portion of a colony of Aspergillus niger in which 
a tan spored mutation appeared as a V-sector : Ai, typical black head of parent strain; 
A 2 , tan head of naturally occurring mutation; X 25 approximately. B and C , Typical 
strain of Aspergillus fumigatus and a naturally occurring mutation discovered and 
described by Edward Yuill as Aspergillus fumigatus mut. helvola. The mutations 
in both species have proved completely stable in continued culture. 



17 C). In the same report a white-spored mutant of A. nidulans, isolated 
in 1937, was described as A. nidulans mut. alba. In both cases the mutants 
developed as natural phenomena without the application of any artificial 
stimuli — in the former case as a single head, in the later case as a group 
of heads, and in both cases from wholly typical strains (fig. 17). 




Plate II 

-Mutations in Aspergillus terreus Thorn produced by irradiating conidia with ultraviolet light. Colonies 
tin >\\ n upon C'zapek's solution agar at room temperature for two weeks. A (upper left), Aspergillus terreus, 
XRRL No. 265, unirradiated stock culture. B (upper right ), Albino mutant, producing white conidial heads 
but retaining the basic cultural and morphological characteristics of the parent culture. C (center left), floc- 
cose, yellow-white mutant producing few and atypical conidial heads. D (center right), Leathery mutant, 
producing tough, close-textured colonies and very few and atypical conidial heads. E (lower left), Nitrate 
mutant, a form unable to use nitrate nitrogen, producing thin spreading colonies and few but entirely typical 
heads. F (lower right), Thiamin mutant, a form unable to produce thiamin, producing thin, spreading 
colonies and very few and atypical conidial heads, see text p. 75. (Color photographs by Haines, Northern 
Regional Research Laboratory. Reproduced through co-operation of Chas. Pfizer & Co., Inc.) 



VARIATION 73 

In studying a group of cultures three years ago, the authors noted a few 
tan heads in the form of a Y-sector in an otherwise typical black colony of 
A. niger (fig. 17A). Isolations from a tan head reproduced the mutant head 
characters, and repeated transfers of this strain have proved consistently 
stable over a period of three years. The mutant strain cannot be 
distinguished from A. niger mut. cinnamomeus (A. cinnamomeus of Schie- 
mann). 

In dealing with natural as well as induced mutations, we have endeavored 
to limit the use of the term mutant to forms whose origin was definitely 
known. Blochwitz (1934, 1935) was not so precise. Under the name 
A. glaucus mut. alba he refers to a white-spored member of the A. glaucus 
group. He believed Aspergillus giganteus Wehmer to represent essentially 
a long-stalked .4. clavatus, hence designated it A. clavatus mut. giganteus. 
This treatment may be justifiable. We believe, however, that until their 
origin from other and well-marked species can be proved, it is wise to con- 
tinue to recognize as species these very distinct forms that are isolated from 
and are able to maintain themselves in nature. 

Cladosarum: The most striking variant, or mutant, ever described in the 
Aspergilli is Cladosarum olivaceum of Yuill and Yuill (1938) . This appeared 
in a culture of A. niger growing on bread at 28° C. (Personal correspond- 
ence). Its colony, conidiophores, vesicles, and primary sterigmata are 
those of Aspergillus. The secondary sterigmata, instead of producing co- 
nidia, thrust out cells which are essentially the same in morphology as the 
secondary sterigmata themselves; the same procedure is then repeated 
several times. Occasionally, however, a terminal cell changes and thrusts 
out several equal cells; in other words, it resumes the function of a primary 
sterigma. The new secondaries repeat the process of producing chains of 
cells each resembling the basal cell with the aspect of a sterigma not a conid- 
ium, and always with the youngest cell at the tip of the chain. In Asper- 
gillus the sterigma which produces a chain of conidia always produces the 
new conidium at the base of the chain, shoving the next most recent farther 
out. Differing then from Yuill 's interpretation, Cladosarum produces no 
conidia, however readily any cell detached from the mass may grow. 

In Aspergillus the ordinary nuclear procedure in conidium formation 
involves mitosis in the sterigma actually producing the conidia. After 
each mitosis one daughter nucleus migrates through the tube into the new 
spore in which it "rests" until that spore begins to germinate. The other 
nucleus remains in the sterigma and repeats the process. This goes on 
until there may be a chain of 200 conidia — the oldest at the outer end, the 
newest directly attached to the sterigma. 

In the absence of cytological study, one may offer the following 
hypothesis. In "Cladosarum" the nuclear procedure must be reversed. 



74 A MANUAL OF THE ASPERGILLI 

The same mitosis occurs. One active and one resting nucleus result, but 
the resting nucleus remains in the sterigma while the active nucleus moves 
into the newly forming cell. This determines the course of development. 
The active, multiplying nucleus is always in the newest cell formed. 

Previously Barnes (1928) had stimulated a strain of the A. glaucus group 
(identified by us as A. amstelodami, 1941) by heat, and reported certain 
mutants which were deposited with Dr. Westerdijk at Baarn. Among 
them, under the designation "Creamy" (NRRL No. 143), a mold with the 
morphology of Cladosarum appeared. It was obviously derived from 
some A. glaucus strain and is, in so far as the writers are aware, the only 
other appearance of the Cladosarum structure ever discovered. Barnes 
does not appear to have recognized its contrasting structure. 

Since no collector has reported this type of mutant in nature, it must 
either be very rare or be unable to maintain itself in a competitive environ- 
ment. However readily such mutants may be maintained in the labora- 
tory, they would rarely reach the second generation in nature on account 
of lack of spores. The name Cladosarum was thus applied to a defective 
organism (zoologically designated a "monster"), which does not become a 
component of any natural flora, hence taxonomically the name should 
be untenable. 

INDUCED VARIATION 

Striking mutations have been obtained from various species of the Asper- 
gilli by subjecting them to artificially imposed stimuli. Schiemann (1912) 
was among the first to draw attention to the possibilities inherent in this 
approach. By subjecting a strain of A. niger to various concentrations of 
potassium bichromate, she was able to produce two striking mutations 
which she designated according to color, A. fuscus (= A. niger mut. schie- 
manni of this manual) and A. cinnamomeus (= A. niger mut. cinnamomeus 
ibid.), respectively. Both cultures have remained stable in our hands, and 
in various collections, throughout the 32 years since their original isolation. 
A third "mutation" designated A. niger var. altipes could not be 
distinguished from other strains of black Aspergilli isolated from nature — 
the parent strain was not seen. Working with a member of the A. glaucus 
group designated Eurotium herbariorum Wigg., Barnes in 1928 reported 
the production of a series of variations by exposing spores to heat. While 
there are reasons for questioning the correctness of some of Barnes' inter- 
pretations and conclusions (see Thorn and Raper, 1941), there is evidence 
that he succeeded in producing a mutant which, in its habit of growth and 
in the character of the fruiting structures developed, bears a striking re- 
semblance to a form subsequently isolated from A. niger by the Yuills 
(1938), and described by them as Cladosarum olivaceum, genus and species 



VARIATION 75 

new. Galloway (1933) obtained marked variation in colonies of Asper- 
gillus terreus by growing them upon media containing flour to which was 
added 0.003 to 0.005 per cent of salicylanilide. 

Thorn and Steinberg (1939), and Steinberg and Thorn (1940a, 1940b) 
in a series of experiments, applied chemical stimulants to a strain of A. 
niger (Thorn No. 4247: NRRL No. 334) which had been in the collection 
many years without noticeable change. From the cultures resulting, 
Steinberg picked out and purified for study all variants he could observe 
with the naked eye and with the aid of a handlens. In examining a 
fruiting area of a colony, general changes were not common; ordinarily an 
occasional head changed color, long or short conidiophores appeared in 
spots, gross malformation showed as areas of no fruit, or too much fruit, 
or color effects in the mycelium. These were picked out and grown in 
successive cultures. Expressed in terms of morphology, the most striking 
feature of the tested culture was disturbance of uniformity. The same 
types of changed aspect were reproduced many times. In general, they 
followed the same lines as have been described as present in the natural 
series selected by Biourge or in the collections of Mosseray at Brussels. 
In general, these changes were destructive in character and included large 
numbers of "injury mutants", or variants, which reverted in subsequent 
transfer to the original aspect of Steinberg's culture. Sodium nitrite was 
the most effective agent used. Some isolations, however, represented clean- 
cut mutations. Strains of A. fumigatus with albino heads were obtained. 
Forms of A. niger with light brown to cinnamon-colored spore heads, es- 
sentially like those earlier obtained by Schiemann (1912), those obtained 
by Whelden (1940), and those subsequently obtained by Raper, Coghill, 
and Hollaender (see below) from members of the same group, were likewise 
isolated. These have remained stable in culture for the four years that 
they have been in our collection. 

Whelden (1940) succeeded in obtaining a series of mutants in A. niger 
by bombarding conidia with low voltage cathode rays and subsequently 
isolating colonies which developed from such irradiated cells. Forms pos- 
sessing heads in various brown shades, rather than black, were isolated, 
as well as one giant form with conidial structures appreciably larger than 
the parent. By means of ultra-violet irradiation of spores, Raper, Coghill, 
and Hollaender (in press) obtained mutants which produced tan-colored 
conidial heads but otherwise closely resembled the parent strain. In a 
more exhaustive study of A. terreus (PI. II, A), the same investigators suc- 
ceeded in isolating a number of striking and markedly different mutations. 
These included albino forms with colorless conidial heads (PI. II, B); forms 
with pale buff-colored heads; yellow, yellow-white, floccose forms with few 
and smaller conidial heads (PI. II, C); forms producing very thin, sparsely 



76 



A MANUAL OF THE ASPERGILLI 



sporing colonies; forms producing restricted colonies characterized by the 
production of an excessive amount of orange-brown exudate; and forms 
with leathery, close-textured colonies bearing very few conidial heads 
(PI. II, D). Alterations in microscopic details commonly accompanied 
these changes in colony appearance (fig. 19). In addition to these morpho- 
logical mutants, which were found to be stable when checked through ten 
successive transfers over a period of 12 months, various physiological 
mutants were also isolated. These included forms unable to utilize nitrate 
nitrogen (PI. II, E) but able to grow and sporulate normally upon media 
containing ammonia nitrogen, and a form unable to synthesize thiamin 
(PI. II, F). Upon Czapek's solution agar containing sodium nitrate and 
sucrose, each of these is strikingly different from the parent strain; upon 




A 




Fig. 18. Induced mutation. A and B, Aspergillus niger group, strain NRRL No. 
67: A, parent culture growing on Czapek's solution agar, 10 clays, room temperature; 
B, tan-spored mutation of same produced by ultraviolet radiation. 



malt extract agar, which contains adequate amino nitrogen and thiamin, 
neither could be differentiated from the parent (Raper, Coghill, and Hol- 
laender, in press). Thus the need for comparative study and examination 
upon a variety of media is apparent, while the importance of such studies 
in reliable taxonomic work cannot be over-emphasized. In cultures of 
Aspergillus terreus resulting from irradiated spores the capacity to produce 
itaconic acid varied from zero in some isolations to levels somewhat above 
the parent culture in others. The majority of such isolations produced 
yields somewhat lower than the parent strain, many produced yields ap- 
proximately equal to it, while a very few produced superior yields (Lock- 
wood, Raper, Moyer, and Coghill, in press). 

Based upon our own investigations and the published reports of other 



VARIATION 



77 



workers, certain observations of a summary character can be made regard- 
ing variation in the Aspergilli: 

1 . Aspergilli include strains and species adapted to a very wide range of 
environmental conditions. Such conditions may influence materially the 
cultural and morphological characteristics of these molds. 






ft ' 




B 



,A 



c 



Fig. 19. Photomicrographs showing details of structure in the conidial heads of 
the parent strain, and in two selected mutations of Aspergillus lerreus (NRRL No. 
265) produced by ultra-violet, radiation, X 600 A, Typical heads of non-irradiated 
parent strain. B, Mutation in which conidium formation is incomplete and cells 
adhere in long chains in liquid mounts. C, Mutation in which many fruiting struc- 
tures develop vesicles but often fail to produce sterigmata and spores. 



2. Under natural conditions, great numbers of variants appear along with 
occasional sharply separable forms, or mutants, which are definitely of 
species rank. The extent to which natural mutations may account for 
described species is a matter of conjecture. 

3. The Aspergilli can be made to mutate in the laboratory by subjecting 
them to a variety of different excitants, or stimuli. Induced mutations may 
parallel some of those found in nature and described as species. 



78 A MANUAL OF THE ASPERGILLI 

4. Particular species of the Aspergilli tend to mutate along certain 
definite lines, e.g., the production in A. niger of forms with tan to light 
brown spore heads (Schiemann, 1912; Steinberg and Thorn, 1940; Whelden, 
1940; and Raper, Coghill, and Hollaender, in press), and the production 
mA.fumigatus of forms with colorless spore heads (Yuill, 1939; and Stein- 
berg and Thorn, 1940a). The type of mutant produced is not governed 
by the type of treatment given, although the number of mutations produced 
is strongly influenced by this factor. 

5. Great variability in biochemical activity is encountered among strains 
isolated from nature, but these differences are rarely linked with specific 
morphological changes. 

6. The Aspergilli can be made to mutate physiologically as well 
as morphologically by the application of various stimuli. Physiological 
mutations may conceivably be of tremendous importance in the develop- 
ment of improved strains for fermentation processes. 

7. Despite natural variation, most strains of Aspergillus when subjected 
to critical transfer and maintenance under rigorous culture conditions can 
be kept for many years with constant colony appearance, stable 
morphology, and dependable biochemical activity. 




PART II 



THE MANUAL PROPER 



Chapter VII 
THE USE OF THE MANUAL 

Since the purpose of this manual is to facilitate the identification of 
Aspergilli as they are isolated from nature and as they are encountered in 
the investigation of special problems, the procedures and considerations 
involved in the use of the manual must be discussed. The general mor- 
phology and structural details found in the spore-producing apparatus of 
the Aspergilli have been described and figured in Chapter III. Com- 
plexities in the specific combinations of these characters found in the exam- 
ination of moldy material and in the isolated colonies of individual strains 
makes desirable a summary outline of the exact observations to be made 
in describing an Aspergillus. Such a descriptive sheet is presented as page 
82. For practical use, a standard sheet of record paper is folded over on 
the left-hand margin for about 5 cm. The column of observations desired 
is written upon this marginal fold ; a fresh sheet of paper is slipped under the 
fold and the descriptive data are filled in, appearing exactly in the same 
order for each strain studied. A single glance at the sheet shows the dis- 
crepancies, if any, in the descriptive data obtained. With such a sheet 
properly filled out, the keys to groups and within groups facilitate the 
placement of the strain in its proper group first, then its allocation to species 
within the group. 

Such descriptive sheets, to have comparative value in species diagnoses 
must present their data in standardized terms. It has, therefore, been 
necessary to define and illustrate the morphological terms accepted in this 
manual, and to indicate as synonyms in the chapters on morphology the 
usages of various describers of Aspergilli back over the 200 years since 
Micheli. 

Identification from specimens : The field mycologist working with speci- 
mens collected and examined fresh or dried will often find completion of a 
technical description very difficult and some observations impossible. One 
with long acquaintance with the Aspergilli may place his specimen to the 
group or aggregate species correctly, but even such workers are frequently 
puzzled. If the organism is deemed important, the fresh or recently col- 
lected specimen should be taken to the culture laboratory to insure its 
isolation and preservation in pure form. The descriptive data at hand 
should then be checked and supplemented from the pure culture. 

To identify an unknown Aspergillus, the worker needs pertinent data 
which will permit him to interpret his mold in terms of species already de- 

81 



82 



A MANUAL OF THE ASPERGILLI 



scribed— including the observations essential in a species characterization. 
For convenience such data may be indicated vertically upon a descriptive 
sheet which is elaborate enough to include observations that are regarded 
as useful. Measurements should be presented as ranges encountered in 
the examination of many units, not as exact and single measurements of 
individual cells or structures. 



Primary sterigmata 

measurements 

arrangement 

color 
Secondary sterigmata 

measurements 
Conidia 

color 

measurements 

markings 
Perithecia 

color 

size 

shape 
Ascospore 

color 

size 

markings 
Sclerotia 

color 

size 



Aspergillus — identifying number or marks. 
Culture medium or natural substratum 
Temperature of incubation 
Colony characters 
Rate of growth 
Texture 
Mycelium 

submerged 
floccose 
color: above 
reverse 
Heads 
color 
form 

measurements 
Conidiophore 
length 
diameter 
wall: thickness 
markings 
color 
Vesicle 
shape 
size 
color 



With such a descriptive sheet before him, the user of the manual finds 
that the Aspergilli have been arranged into a series of natural groups (fig. 
20), each containing one to several species aggregates. Each group in- 
cludes species with varieties, and at times mutants, having a series of es- 
sential characters in common. These groups have been arranged as nearly 
as possible in natural order, based upon the presence or absence of certain 
contrasting intergroup characters. 

These major separating characters are usually evident and positive. 
Nevertheless, individual species are found in which certain of these charac- 
ters are reduced to vestigial or apparently suppressed, yet which show so 
many characters allying them with a particular group that such placement 
is more logical than any other. Such species must sometimes be arbitrarily 
placed, and their possible affiliation with other groups indicated both in the 
discussion of the species and in the discussion of the related group. 



THE USE OF THE MANUAL 83 

Species 

The species concept in Aspergillus is very difficult to define in tangible 
terras. In this manual, the species names already in use have been pre- 
served wherever possible. The actual material originally described under 
a particular species name (i.e., type material) exists for but a few species. 
If such material exists, it is more important as fixing one point, one individ- 
ual strain in a series of intimately related variants, than tying the name 
to extremely definite morphology. If such material is not known, compari- 
son of large numbers of strains in pure culture with authoritative descriptive 
information, supplemented by laboratory usages coming down from the 
original describer, usually fixes the series or form intended. 

From such composite sources it is commonly possible to establish a fairly 
concrete morphological aspect based upon ranges of color, differences in 
structure, and variations in spore measurement which are repeated in great 
numbers of isolates. In such series of isolates, there are no sharp lines of 
demarcation when large numbers of strains are brought together. Within 
our concept, a single strain may show much of this variability within its 
colonies in culture, or it may reduce or suppress certain characteristics and 
intensify others. Such variants have often been given species rank by 
workers unaware of the existence of other variants completely bridging 
the gap between such forms and other members of the series. Great dif- 
ferences in biochemical activity may be shown by different strains with or 
without contrasting morphology. Nomenclature based upon an assumed 
correlation of a particular cultural aspect with industrial significance has 
been offered but has proved utterly unreliable in identifying an organism 
if lost, or in seeking a new strain to serve the same purpose. 

Two contrasting tendencies in classification are always encountered. 
In the Aspergilli these may be represented by Mosseray (1934a) who found 
diagnostic marks to distinguish 35 species among 63 cultures of black 
Aspergilli in the collection of Biourge at Louvain. He later received many 
more variants and faced the question whether to try to describe them all or 
abandon the field. He admitted inability to write descriptions explicit 
enough to identify them all. In contrast, Neill (1939), disregarding asco- 
spore measurements and markings, "lumped" all of the A. glaucus group 
into A. glaucus Link. Likewise, all of the black Aspergilli were considered 
as A. niger van Tieghem. Forms that he did not happen to recognize as 
belonging to one of his groups were discarded. These are extremes. 

With abundant living material before him, the student of the Aspergilli 
can usually recognize as representative a reasonable number of forms which 
can be described in tangible specific terms. Commonly, forms which actu- 
ally play a significant role in nature or in biochemical processes can be 
selected as the points around which such species descriptions are drawn. 



84 A MANUAL OF THE ASPERGILLI 

On the other hand, forms such as A.janus, A. itaconicus, A. lutescens, etc., 
while probably rare in nature, possess sufficiently distinctive morphology 
to warrant species recognition irrespective of other considerations. 

Varieties 

The taxonomic term, variety, is used here to designate any homogeneous 
member of a species complex which carries most of the diagnostic characters 
of the species but maintains one or more clearly defined differences in 
particular characters. For example, variety alba is used for certain strains 
of particular species in which the characteristic color of that species is 
absent. 

There is little agreement in the literature in the application of the term, 
variety; certain authors use the term to indicate their belief that one form 
with particular morphological characters had its origin from another. 
In such cases, the belief is hypothetical, not a matter of observation. 
Sometimes previously known species were merely moved to varietal stand- 
ing without specifying the characters upon which the decision was based. 
Such changes are reduced to synonymy or, if entirely unsupported, are 
occasionally ignored in this manual. The term variety is only useful if 
definitely associated with a clearly defined variation in structures within an 
otherwise homogeneous series of strains. 

Mutations, or Mutants 

The term mutation, or mutant, is only recognized here for forms resulting 
from a sharp break in morphology (including color) from known structures 
characteristic of a species, to a definitely altered and inherited contrasting 
structure. Obviously the only excuse for the term in taxonomic usage is to 
designate the origin of the form studied. If the source of such a variant 
were unknown, the taxonomist would designate the form present as a 
variety or species, depending upon the nature and importance of the changes 
encountered. The increasing number of studies in experimental evolution 
make recognition of induced variation taxonomically necessary. 

New Species 

The discriminating collector will occasionally find an organism markedly 
divergent in characters from any described form. Usually these diver- 
gences leave the organism readily recognized as a member of one of the great 
groups. If the differences in aspect and detail of structure separate such 
a form from the other described members of the group, and if the form is 
found often enough to prove that it has a place in nature, description as a 
new species is warranted. Similarly, an occasional form, either by sup- 



THE USE OF THE MANUAL 85 

pression or complete disappearance, loses the arbitrary diagnostic character 
which furnishes the basis for separating two adjacent groups. In such cases 
it has at times seemed more practical to add such a species to the group most 
nearly allied to it by general colony aspect, but to cross-reference it to the 
related group. 

The detection and description of species hitherto unrecognized neces- 
sitates extensive review of the literature and restudy of available living 
cultures of at least one whole section of the great genus Aspergillus. Unless 
the one who encounters a form that he cannot recognize under names 
already in the literature is prepared to investigate his form adequately in 
relation to the whole genus, he should not describe his organism as a new 
species. 

Summarized 

Comparative study of the taxonomic literature brings out the need for a 
standardized series of morphological and descriptive terms into which the 
many usages introduced in the two centuries since Micheli can be translated. 
Variations in measurement are deemed significant only if they exceed the 
common limits between closely related organisms and predominate in the 
preparations examined. Ranges in measurements are more significant 
than exact dimensions of either selected structures or averaged values based 
upon many measurements. 

Merely quantitative variations are not recognized as warranting separa- 
tion of species. For example, differences in the shade of color, or the in- 
tensity of a particular reaction, especially when other strains are found 
to fall between the "old" and proposed "new" species, are not regarded as 
species characters. Such proposed names either fall to varietal status or to 
synonymy. 

The names not accepted here fall into several categories. (1) Many are 
listed as synonyms because they are believed to have been given to variants 
not recognizable by dependable and interpretable differences from other 
members of the same series. (2) Fantastic variants, or "monsters", 
appearing in culture may, like "Cladosarum", be maintained in the labora- 
tory, but unless found perpetuating themselves in nature, clearly fall in the 
class of "natures experiments" which do not contribute to the permanent 
flora. Such names are not regarded as established. (3) Unidentifiable 
species — names appearing in the literature based upon structures or re- 
actions regarded by the describer as unique, but whose identity is so com- 
pletely lost in large collections among series of closely related strains as to 
make them unidentifiable by description — are listed in the check list with- 
out characterization. 



86 A MANUAL OF THE ASPERGILLI 

SPECIES KEYS 

No general or comprehensive key to the species of Aspergillus is pre- 
sented. Instead, a series of comparatively simple species keys are included 
in the discussions of the several groups. The recommended procedure in 
identifying an Aspergillus with the aid of this manual is first to determine 
its group relationship by means of one or more of the group keys presented 
below, then assign the culture more precisely to species by means of the 
intra-group species key for the particular group to which the form belongs. 

GROUP KEYS 

Three keys to the groups of Aspergilli are offered : The first is presented 
in the form of a diagram and presents the different groups in what we con- 
sider their natural order. Presentation of this key in graphic form, it is 
believed, will materially assist the user in grasping the various characters 
which ally and interrelate the different groups. Primary separation is 
based upon the number of series of sterigmata, whether single or double. 
Secondary separation is based upon the character of the conidiophore, 
whether rough or smooth. Tertiary separations are based upon the 
presence or absence of perithecia, hulle cells, and sclerotia, and upon 
the color of the conidiophore wall. 

In assigning species to groups by means of this key, the transitional or 
intermediate character of certain species becomes strikingly apparent. For 
example, Aspergillus caespitosus possesses the brown conidiophore and 
conidial coloration of A. nidulans, furthermore, it produces clusters of ir- 
regular, thick-walled hulle cells; but the head is radiate, or only loosely 
columnar, and no perithecia or ascospores are produced. It is placed in the 
A. nidulans group with full recognition that it possesses certain characters 
which relate it to A. ustus. Aspergillus alliaceus is another form with 
intermediate characters. The conidiophore is uncolored and smooth when 
examined in liquid mounts (appearing finely roughened when examined 
dry), and the sclerotia are black, but the heads are essentially ochraceous 
in color. It is placed in the A. wentii group but shows unmistakable re- 
lationship to A. ochraceus. Aspergillus sparsus is a species of uncertain 
relationship. It possesses a conspicuously roughened, yellow conidiophore 
and globose head, and is placed in the A. ochraceus group; but the conidial 
heads show a greenish color which is not found in any other known member 
of this group, while the character of the conidiophore will not warrant place- 
ment elsewhere. The so-called "bronze series" in the A. tamarii group 
is transitional in the direction of A. flavus. Colonies are conspicuously 
green when young and retain a greenish tint for a considerable period, in 
contrast to A . tamarii which never shows true green and appears greenish 
only transiently when young. Such a list of intermediate species and forms 



THE USE OF THE MANUAL 



87 



A.clavotus Group 

(Heads clovote) 




\ 


_ 


.J 




A.glaucus Group 

(Penthecia yellow) 


> 


>Ascosponc 


^ Single 
x Stengmato 


A.fumigatus Group 




A.nidulans Group 

(Ascospores red) 


~"\ 


\ 


-> 




V Sclerotia 
r Laciung 


^Conidiophores 
f Smooth 




A.ustus Group 


IConidiophore 








A.flavipes Group 


J 


Brownish 


V huiie 
/ Cells 

J 




A. versicolor Group 






A.terreus Group 


J 




A.candidus Group 


> 




A. niger Group 

(Heads globose, in "block" shades) 


V Double 

r Slengmata 


A. wentii Group 










A. tamarii Group 




VSclerotia 
r Present 


VConidiophores 
/^ Rough 

> 




A.flavus-oryzae Group 

(Heads yellow— green) 








A.ochraceus Group 

(Heads ochre to yellow) 


> 


Comdiophore 
> Walls 
Yellowish 




; 





Fig. 20. Graphic representation of natural relationships among groups comprising 
the genus Aspergillus. The abundance of species in the different groups is roughly 
indicated by the size of the group boxes in the figure. 



could be extended, but sufficient have been cited to indicate that the various 
groups, like the species which comprise them, often cannot be set apart 
by sharp lines of demarcation. 



88 A MANUAL OF THE ASPERGILLI 

In using this manual, and in studying the Aspergilli generally, it is im- 
portant that the worker should realize that these organisms vary within 
the species, the species vary within the group, and to a lesser extent, the 
groups themselves vary within the genus. In other words, nature did 
not realize that we were going to write this manual when the various 
species and groups were being developed, hence not all of the forms one 
encounters will fit into the various compartments which have been con- 
structed, although these are, on the whole, comparatively elastic. This can 
be illustrated in another way. If we completely disregard color, the genus 
Aspergillus, as depicted in figure 20, can be likened to the spectrum. There 
is a green region and a yellow region in the spectrum and the two regions are, 
on the whole, distinct. Furthermore, within each of these, certain fixed 
and definite lines can be identified. It is, however, extremely difficult, if 
not impossible, to say where the green region ceases and the yellow begins. 
So it is with the species and groups of the Aspergilli. There is a definite 
nidulans group and a definite ustus group, but the line separating the two is 
extremely tenuous. 

We do not, in any sense, infer that the Aspergilli cannot be classified — 
that they cannot be separated into groups, species, and even varieties. 
This manual is evidence that they can. But we do wish to emphasize that 
we are dealing with living and variable organisms, and that in describing 
them, we should be as explicit as possible and still keep our concepts reason- 
ably elastic. 

Key to Groups — Based Primarily Upon Color 

The second key is based primarily upon color and is entirely artificial 
in its construction. The various groups are separated by contrasting 
coloration, and closely related groups may appear widely separated in the 
key. In practice, such a key is very useful since color is the most obvious 
character of an Aspergillus, and since the species comprising a particular 
group, with but few exceptions, are characterized by variations in shade of 
color rather than differences in basic coloration. Presumptive assignment 
of the Aspergilli to groups can usually be made from this type of key which 
is based primarily on color supplemented by the use of a handlens or dis- 
secting microscope. 

A. Conidial heads in definitely green, blue-green, or yellow- 

green shades in young fruiting colonies B. 

AA. Conidial heads lacking green colors (Greenish in excep- 
tional cases) K. 

B. Conidial heads in green and blue-green shades C. 

BB. Conidial heads in yellow-green shades A. flavus group 



THE USE OF THE MANUAL 89 

C. Conidial stalks and heads coarse— heads clavate A. clavatus group 

CC. Heads not clavate D. 

D. Colonies mostly showing yellow perithecia and more or 

less yellow and red hyphae A. glaucus group 

,DD. Colonies lacking yellow perithecia and more or less yel- 
low and red hyphae E. 

E. Colonies producing columnar spore masses F. 

EE. Colonies producing radiate, globose, or hemispherical 

heads H. 

F. Rapidly growing and spreading colonies G. 

FF. Slowly and restrictedly growing colonies A. restrictus series 

G. Conidial columns long, narrow A.fumigatus group 

GG. Conidial columns short and broad; perithecia usually 

present, ascospores red A. nidulans group 

H. Heads radiate in blue-green, dull green, to pale tan or 

flesh-colored shades A. versicolor group 

HH. Heads in some other color L. 

K. Heads in long compact columns, avellaneous to cinna- 
mon, shading toward colorless through light flesh 
colors A. terreus group 

KK. Heads in some other color L. 

L. Colonies more or less floccose; heads in dull olive-grays 

to fuscous A. ustus group 

LL. Heads in some other color M. 

M. Young heads white or only slightly tinged in age N. 

MM. Heads in some other color O. 

N. Young heads white, usually in short columns, broad- 
ening at apex, often becoming avellaneous in age. . . . A. flavipes group 

NN. Heads persistently white, larger heads definitely globose 

or radiate A. candidus group 

O. Heads in sulphur yellow to ochre shades A. ochraceus group 

00. Heads in some other color P. 

P. Young colonies showing a greenish color passing into 

brown A . tamarii group 

PP. Heads not showing greenish Q. 

Q. Heads in purple-brown to black shades A. niger group 

QQ. Heads in yellowish-brown shades, orange to deep brown 

to umber color A. wenlii group 



90 A MANUAL OF THE ASPERGILLI 

Key to Groups — Based Primarily Upon Morphology 

The third key is based primarily upon morphology, with colony color 
employed as an accessory differentiating character. This key is also arti- 
ficial in construction and is designed to separate the various groups by the 
simplest and most direct means possible. Groups naturally related may 
or may not appear in their proper sequence. With the data developed 
upon the descriptive sheet (p. 82), most of the Aspergilli can be traced 
to their proper placement in classification schemes by using the following 
key. Natural arrangement is disregarded and the same group is occasion- 
ally reached in different places in the key. 

A. Species producing perithecia and ascospores B. 

AA. Species not producing perithecia and ascospores D. 

B. Ascospores colorless C. 

BB. Ascospores purple-red A. nidulans group 

C. Perithecia white to flesh color, enmeshed in a loose net- 

work of colorless hyphae A . fischeri 

CC. Perithecia yellow to orange, naked, vegetative hyphae 

often showing red to orange granules A. glaucus group 

D. Conidial heads cylindrical -clavate; vesicles definitely 

clavate A. clavalus group 

DD. Conidial heads not cylindrical -clavate E. 

E. Colonies showing green or greenish color at some stages 

of development F. 

EE. Colonies lacking green color P- 

F. Conidiophore wall rough or pitted G. 

FF. Conidiophore wall smooth H. 

G. Colonies green or yellow-green to yellowish A . flavus-oryzae group 

GG. Colonies greenish-brown when young, becoming rich 

brown or umber in age A. lamarii group 

H. Sterigmata in one series I- 

HH. Sterigmata in two series K. 

I. Conidia elliptical to pyriform J- 

II. Conidia spinulose, 2.5 to 4 M , globose; chains in com- 

pact columns A. fumigatus group 

J. Colonies mostly showing yellow perithecia; sterigmata 

usually coarse A. glaucus group 

J J. Colonies lacking perithecia; conidia in narrow columns. A. reslrictus group 



THE USE OF THE MANUAL 91 

K. Conidiophores in yellow-brown shades L. 

KK. Conidiophores not colored 0. 

L. Conidial heads definitely green M. 

LL. Conidial heads greenish only when young, then in yel- 
low-brown shades A . ustus 

M. Ascospores produced— purple-red in color A. nidulans group 

MM. Ascospores not produced N. 

N. Colonies showing irregularly clustered hiille cells A. caespitosus 

NN. Colonies not showing hulle cells: bright green, spreading A. unguis 

O. Conidial area in dull green shades (sometimes partially 

or completely replaced by tan) A. versicolor 

00. Conidial area in blue-greens A. sydowi 

P. Conidiophore walls smooth Q. 

PP. Conidiophore walls rough V. 

Q. Conidiophore walls pale yellow in outer layer; heads 

white when young often becoming avellaneous in age. A. flavipes 

QQ. Conidiophore walls colorless, or partially yellow-brown 

near the head R. 

R. Conidial chains in solid columns, compact at base S. 

RR. Conidial chains radiate at least in the larger and typical 

heads T. 

S. Conidial heads in avellaneous shades A. terreus 

SS. Conidial heads flesh color (in pinkish shades) A. carneus 

T. Heads white or tardily in yellowish shades A. candidus group 

TT Heads in darker colors U. 

U. Conidial heads globose in purple-brown to black, (rarely 

brown or paler) A . niger group 

UU. Conidial heads globose in yellowish, yellow-brown, and 

dark brown shades A . wentii group 

V. Conidiophore walls rough, yellow; heads yellow to 

ochre A . ochraceus group 



Chapter VIII 
THE ASPERGILLUS CLAVATUS GROUP 

Outstanding Characters 

Conidial heads clavate 1 , large, pale blue-green. 
Conidiophores generally coarse, smooth-walled, uncolored. 
Sterigmata in one series. 
Conidia elliptical, smooth, comparatively thick-walled. 

Group Key 

Conidial structures not exceeding 4.0 mm. in length. 

Aspergillus clavatus Desm. 
Conidial structures often 1 to 5 or more cm. in length. 

Aspergillus giganteus Wehmer. 

Aspergillus clavatus Desmazieres, in Ann. Sci. Nat. Bot. (2) 2: 71, p. 2, 

fig. 4. 1834. 

Colonies upon Czapek's solution agar growing rapidly at 20-24° C, 
plane or slightly furrowed, in certain strains tending to become floccose 
but generally characterized by a surface mycelial mat and abundant erect 
conidiophores up to 3.0 mm. in length, bearing large, blue-green, clavate 
conidial heads evenly distributed or arranged in more or less well defined 
zones (PI. Ill, A and Fig. 21 A); reverse generally uncolored, but becoming 
browned in age in some strains; odor strongly foetid in some strains, not 
pronounced in others. Conidial heads clavate, large, commonly ranging 
from 300 to 400m by 150 to 200m, in age splitting into 2, 3, or more divergent 
columns of compacted conidial chains (Fig. 21 B), approximately slate- 
olive in color (Ridgway, PI. XLVII). Conidiophores 1.5-3.0 mm. in 
length, 20 to 30m in diameter, comparatively thin-walled, smooth, color- 
less, gradually enlarging at the apex into a clavate vesicle which is fertile 
over an area up to 200 to 250m in length and 40 to 60m or more wide (Fig. 
21 C). Sterigmata in a single series, varying in size from 2.5 to 3.5m by 
2.0 to 3.0m at the base of the vesicle to 7.0 or 8.0 and occasionally 10m by 
2.5 to 3.0m at its apex (Fig. 21 D). Conidia elliptical, comparatively 

1 Aspergillus janus Raper and Thorn (see page 187) is characterized by a smaller 
clavate vesicle in one of its conidial phases. It is hardly to be confused with the 
clavatus group, however, because of the whiteness of its conidial masses, the double 
series of sterigmata, and the intermixture of more or less abundant green A. sydowi- 
like heads in cultures at room temperature. 

92 



THE ASPERGILLUS CLAVATUS GROUP 



93 



heavy walled, smooth, 3.0 to 4.0/x by 2.0 to 3.0/j, ocasionally larger in some 
strains and irregular in others. 




D 

Fig. 21. Aspergillus clavatus Desm. A, Colony growing on Czapek's solution 
agar, 10 days, room temperature, X 1.3 (Thorn No. 5169). B, Conidial heads showing 
characteristic splitting in age, X 12 (Strain NRRL No. 1823). C, Conidial heads 
showing characteristic clavate form of vesicle, X 180 (Strain NRRL No. 6). D, 
Portion of a conidial head further enlarged, showing closely packed single series of 
sterigmata, X 600 (Strain NRRL No. 6). 



Cosmopolitan in distribution and especially common in soil, decaying 
vegetation, dung and other materials where active decomposition of nitrog- 
enous materials is taking place. Cultures examined include numerous 



94 



A MANUAL OF THE ASPERGILLI 



strains from various parts of the United States, together with isolations 
from China, British Guiana, Cuba, Panama and European sources. 

The species description as presented is based upon a large number of 
closely related strains that have been examined, and is adequately repre- 
sented by such specific strains as NRRL Nos. 2, 5, 8, and others. 

Upon malt extract agar, details of morphology and colony characteristics 
may or may not conform with those listed above. For example, in such 
typical strains as Nos. 2, 5, and 8, conidial structures are generally more 
abundant upon malt than Czapek's agar and may average as much as 20 to 
25 percent larger in size. In other strains such as Nos. 4 and 6, a markedly 
different response is noted upon this medium. Conidial heads, although 



Q 




// 



'& 







Fig. 22. Aspergillus clavatus Desm. (Strain NRRL No. 1: Thorn No. 107). A, 
Colony growing upon Czapek's solution agar, 10 days, room temperature, X 1.3. 
B, Conidial heads diminutive and somewhat atypical but showing characteristic 
clavate form, X 600. 



greatly increased in number, are much reduced in size, with conidiophores 
generally 1 mm. or less in length bearing heads only 100 to 125/x long and 
proportionately reduced in diameter. In these latter forms, conidia are 
somewhat irregular in form and generally larger than in the more typical 
strains first considered. 

Culture NRRL No. 1 (Thorn No. 107) differs markedly from the species 
description in producing deeply floccose colonies (Fig. 22 A) and compara- 
tively few spore heads which are extremely variable in size. These range 
from very small fruiting structures (Fig. 22 B) borne as branches upon 
aerial hyphae to structures arising from the substratum which are charac- 
terized by dimensions almost typical of the species. This culture is more 
nearly normal upon malt than upon Czapek's solution agar but is unique 



THE ASPERGILLUS CLAVATUS GROUP 95 

among the strains examined by us and must be considered somewhat 
atypical. It deserves particular attention because it was obtained from 
the Centraal bureau in Baarn in 1908 as Aspergillus clavatus Desm. and has 
undoubtedly been widely distributed by that organization — in fact, it is 
quite probable that it has been examined by more investigators than any 
other strain belonging to this group. We refrain from using it as a basis 
for the species description, however, despite its classic history, since the less 
floccose and more heavily sporing strains are so much more commonly 
encountered in nature. 

Members of this species produce a strong alkaline reaction upon many 
culture media and this is usually associated with a strong foetid odor. For 
example, when grown upon Czapek's solution agar containing only NaN0 3 
as nitrogen and sucrose as a carbon source, the reaction of typical strains 
may reach pH 9.5 or even higher accompanied by a strong odor of tri- 
methylamine almost approaching putridity. No other species of Asper- 
gillus is known to react in this manner although some strains of A . flavipes 
give some suggestion of it. The ability to produce, and more particularly 
to withstand strong alkaline conditions undoubtedly accounts for the 
common occurrence of this species upon dung and other nitrogen rich 
substrata undergoing decomposition. 

Aspergillus giganteus Wehmer, in Central, f. Bakt., etc., 2, Abt. 18, No. 

13/15: 385. 1907. 

Colonies upon Czapek's solution agar growing rapidly at 20° C, charac- 
terized by an extensive surface and submerged vegetative mycelium and an 
early development of abundant conidiophores 2.0 to 4.0 mm. high, followed 
by the subsequent development of less numerous conidiophores ranging up 
to several centimeters in length (PI. Ill, B and Fig. 23 A), the latter 
strongly phototropic and generally more abundant in marginal areas, com- 
monly obscuring the more central mass of short conidiophores ; colonies at 
first white, becoming pale blue-green as conidial heads mature ; reverse dull 
tan, becoming brown in age; odor none to somewhat foetid in certain 
strains. Conidial structures varying greatly in dimensions and falling for 
the most part into two general size ranges: (1) conidiophores commonly 
2 to 3 mm., rarely exceeding 4 mm. in height, bearing clavate heads 200 to 
350/x in length; (2) conidiophores one to several centimeters in length, 
bearing heads up to 1 mm. in length. The relative proportions of these 
head types is strongly influenced by environmental conditions, and specific 
strain characteristics. Conidial heads pale blue-green, in age splitting into 
2 or more columns extending the length of the vesicle (Fig. 23 B). Vesicles 
consisting of the expanded terminus of the conidiophore, ranging from 100 
to 250m by 30 to 50m upon short conidiophores to 400 to 600m by 120 to 



96 



A MANUAL OF THE ASPERGILLI 



180/x upon long conidiophores (Fig. 23 C). Sterigmata in a single series 
ranging from 3.0 to 4.0m by 2.5 to 3.0m at the base of the vesicle to 6.0 to 




- . •. ■ . «.tJ» *t •* »* 2 . . *• * 



- .V » 



»■-.*. . i! i •• ' . . "■ ,«» 7 • ;, , 




Fig. 23. Aspergillus giganteus Wehmer (Strain NRRL No. 10). A, Portion of 
colony on Czapek's solution agar showing characteristic long conidiophores and 
large clavate heads; 10 days, room temperature, X 1.3. B, Portion of colony margin 
somewhat enlarged, X 11. C, Single conidial head showing the very elongate vesicle 
characteristic of the species, X 130. D, Terminal portion of vesicle showing the 
closely crowded single series of sterigmata, X 600. 



8.5m by 2.8 to 3.5m at the apex (Fig. 23 D). 
walled, smooth, 3.5 to 4.5m by 2.4 to 3.0m- 



Conidia elliptical, thick- 




Plate III 

-v-dd ' u '<l' er ln ft !' As ? e r(,iUus rIaralus Desm., XRRL No. 8. fi i upper right), Aspergillus giganleus Wehmer, 
id x-Diiv i nciu ? ated at 20 ° C. in one-sided illumination. C (center left). Aspergillus repens (Cda.) 

detfary, NKKL No.20. D (center right ), Aspergillus ruber Bremer, XRRL No. 54. E (lower left), Aspergil- 
lus anstelodam, L/Mang.) Thorn and Church, XRRL No. 90. /•' lower right), Aspergillus niveo-glaucus Thorn 
and Kaper, M(KL .No. 12, . Figures A and B growing upon standard Czapek's solution agar with 3 percent 
sucrose; t to /• , growing upon Czapek's solution agar with 20 percent sucrose. (Color photographs by Haines 
-Northern Kegional Research Laboratory. Reproduced through co-operation of Chas. Pfizer & Co, Enc 



THE ASPERGILLUS CLAVATUS GROUP 97 

The above species description is centered upon strain NRRL No. 10 
(Thorn No. 5581. 13A) isolated from Yucatan caves by Prof. F. A. Wolf 
(1938). Additional strains examined include isolations from Texas, Illi- 
nois, Mexico, Puerto Rico, and strain NRRL No. 1725 (Thorn No. 138) 
received from Dr. Westerdijk in 1910 as A. giganteus Wehmer. 

In the present treatment we include under the species name A . giganteus 
Wehmer all strains which produce conidiophores in excess of 1 cm. in length. 
While this may appear somewhat arbitrary, it is done since members of the 
A. clavatus group seem to fall into two natural series: (1) those which never 
produce conidiophores in excess of 5 to 6 mm. in length irrespective of 
environmental conditions or medium composition, and (2) those which 
regularly produce few to many very long-stalked fruiting structures under 
the usual conditions of laboratory cultivation and examination. To the 
first of these series is applied the species designation A. clavatus, to the 
second, A. giganteus. 

Strains of A. giganteus, like those of A. clavatus differ materially in their 
growth and cultural appearances upon different culture media. More 
striking, however, is their response to light and temperature. This has 
been observed by Wehmer (1907), Wolf (1938) and others, and has been 
studied somewhat exhaustively by Webb (1942). Using the Wolf isolate, 
Webb found that the production of long conidiophores was favored by 
cultivation upon media containing from 1 to 10 percent sucrose and in- 
cubation at 20° C. in the presence of light or darkness, whereas heavy 
conidial production and the development of short conidiophores were 
favored by incubation at 30° C. in darkness. Different strains vary ma- 
terially in their cultural appearance upon such standard media as Czapek's 
solution agar and can be roughly grouped into three sub-series as follows: 
(1) wholly typical strains consistently producing the cultural picture as 
defined for the species, (2) strains producing an unusually heavy crop of 
short-stalked conidial structures in colony centers, followed by the produc- 
tion in marginal areas only of long-stalked fruits typical of A. giganteus, 
and (3) rather sparsely growing strains in which there is a general admixture 
of short-stalked and scattered long-stalked fruits ranging up to 2 to 3 cm. 
in length. The latter group is considered as possibly representing atypical 
and somewhat depauperate strains of the first. The second seems to con- 
stitute a consistent and fairly well-defined cultural entity, but does not 
differ from typical forms sufficiently to warrant separation as a variety. 

The validity of the species A. giganteus has been questioned by some 
authors. Blochwitz (1929) regarded it as a mutation of A. clavatus and 
so designated it in his monograph of the Aspergilli. His view may be 
correct. It is our belief, however, that the species should be retained since 
forms producing the giant conidiophores noted by Wehmer are repeatedly 



98 A MANUAL OF THE ASPERGILLI 

isolated from nature, and since no evidence has been presented indicating 
that these larger forms arise directly from the smaller and more common 
forms. While it is true that conditions can be altered so that A. giganteus 
cultures suggest A . clavatus, no one has yet demonstrated that the reverse 
can be accomplished. 

Group Synonyms 

The following names have been proposed for specimens belonging to this 
group but without adequate data to warrant recognition as valid species : 

A. clavellus Peck, in N. Y. State Mus. Nat. Hist. Rept. 34: 49, PI. 2, figs. 1-5. 
1881. Described from cooked squash in New York State. No data is presented which 
would warrant separation of this form from A. clavatus Desm. 

A. westendorpii Sacc. and March, in Rev. Mycologique 7: 149, 1885, was listed 
from cow dung. Correctly assigned to A. clavatus by Lindau, in Deutsch. Krypt. 
Fl. Pilze 8: 152. 1907. 

A. fusco-cinereus Ellis and Morgan was the name attached to Morgan's jacket 
No. 674, showing a very small, clavate aspergillus which has not been collected again, 
hence never cultivated. It was probably some member of this group. 

A. pseudo-clavatus Purjewitch, in Schrift. Naturforsch. Gesell. Kiev 16 (2): 309, 
pi. 12, 1900; see also Sacc. Syll. 16: 1028. The organism in culture was reported as 
having both primary and secondary sterigmata in a small-sized "clavatus" type of 
head, and perithecia with ascospores which were not adequately described. Until 
somebody finds this organism again and reports its cultivation and more complete 
description, it will remain doubtful. (See Thorn and Church, The Aspergilli, 
p. 100. 1926.) 

Occurrence and Economic Importance 

Members of the A . clavatus group are quite common in soils and decom- 
posing materials characterized by a comparatively high nitrogen content. 
They appear to be common upon the dung of various animals and in the 
writers' experience have been isolated repeatedly from that of chickens. 
While the subject has not been adequately investigated, it is probable that 
the ability of members of this group to withstand strongly alkaline condi- 
tions enables them to operate successfully as agents of decomposition in 
situations where almost all other fungi are eliminated. 

Antibiosis 

Certain strains of A . clavatus produce substances in the substratum which 
are capable of destroying Staphylococcus and other microorganisms. Such 
activity was first reported by Weisner in March 1942 (Nature 149: p. 356) 
and subsequently by Waksman, Horning, and Spencer in August of the 
same year (Science 96: p. 202). To the active substance the latter investi- 
gators assigned the name clavacin and noted that it appeared similar to, if 
not identical with, that studied by Weisner (1942) to which the designation 



THE ASPERGILLUS CLAVATUS GROUP 99 

clavatin has since been applied. Additional studies by Waksman and his 
co-workers have further defined its action and enlarged the list of bacterial 
species inhibited (1942b and 1943). Hooper et al (1944) have demon- 
strated the identity of clavacin and patulin, a bactericidal substance ob- 
tained from Penicillium palulum by Raistrick and associates (1943) and 
reported to be of value in the treatment of the common cold. 

Philpot (1943) reported the production of a penicillin -like substance by 
a strain of Aspergillus giganteus. In earlier tests by Wilkins and Harris 
(1942) this strain had been found to produce a substance active against 
Staphylococci. 



Chapter IX 
THE ASPERGILLUS GLAUCUS GROUP 1 

Outstanding Characters 

Perithecia generally present; yellow, globose to subglobose, thin-walled, 
suspended in networks of red or yellow hyphae. 
Asci 8-spored, without definite arrangement; usually ripening in 2 to 

4 weeks. 
Ascospores lenticular, smooth or rough-walled, generally showing an 
equatorial line or furrow with or without flanking ridges or crests. 
Conidial heads more or less abundant, radiate to somewhat columnar, 
typically in some shade of green. 
Conidiophores smooth-walled, terminating in dome-like vesicles. 
Sterigmata in one series, rather coarse. 

Conidia elliptical to subglobose, uniformly and characteristically 
roughened. 

General Considerations 

Aerial hyphae encrusted with yellow, orange, or red granules are abun- 
dant in perithecial areas of most of the strains of the group. Both labora- 
tory cultures and naturally moldy specimens frequently show this as their 
most conspicuous character, one which is readily recognized with the hand 
lens. In nature, molds of this group appear as patches of green, yellow, 
reddish, or reddish-yellow mold, depending upon the relative abundance of 
conidial heads, perithecia, and encrusted aerial hyphae, and especially 
influenced by the composition of the substratum. 

Representatives of the A. glaucus group are universally distributed in 
nature and are significant in the incipient spoilage of many organic ma- 
terials useful to man. They occur particularly upon products characterized 
by a high osmotic tension such as preserves, jams, cured meats, leather 
goods, improperly dried hay, moist grain, and soft woods stored under 
humid conditions. The classic habitat is improperly dried herbarium 
specimens. 

The earliest references to any Aspergilli concern representatives of this 
group, for botanists early encountered them upon herbarium material. 
Micheli in 1729 used the generic name Aspergillus (rough head) for the 
conidial heads, characterized by divergent chains of spores, commonly 
present upon such specimens. Later in the century, Wiggers (1780) pro- 

1 Abridged from: Charles Thorn and Kenneth B. Raper, The Aspergillus Glaucus 
Group, U. S. Dept. of Agr., Misc. Pub. No. 426, Washington, D. C, September 1941. 

100 



THE ASPERGILLUS GLAUCUS GROUP 101 

posed the name Mucor herbariorum for the yellow perithecia found mixed 
with the Aspergillus heads, which he regarded as a different mold. In 
1809, Link designated the green heads Aspergillus glaucus and the yellow 
perithecia Eurotium herbariorum. Half a century later, DeBary (1854) 
proved that the Aspergillus heads and Eurotium perithecia were borne 
upon the same mycelium, hence were one fungus. Although it could be 
maintained that the name Eurotium (designating the perfect stage) should 
take precedence over Aspergillus (descriptive of the conidial apparatus), 
most recent authors have tended to go back to Micheli and use the name 
Aspergillus for the whole group because of the obvious relationship of many 
conidial forms for which no perithecia are known. 

Laboratory Cultivation 

The pattern and size of the ascospore, when present, is especially signifi- 
cant in describing species of the Aspergillus glaucus group. Nevertheless, 
the conidial apparatus and the vegetative mycelium of particular subgroups 
are so important that pure culture under known conditions is always de- 
sirable. The character of the colony, as well as the amount of growth, is 
strongly influenced by the culture medium, and it is only upon substrata 
characterized by a high osmotic tension that typical perithecia and conidial 
heads are produced. It should be noted, however, that characteristic 
heads and perithecia normally develop, although few in number, in situa- 
tions where less concentrated media dry out rapidly, as at the edge of an 
agar slant. Colony comparisons for correct identification can best be 
made in Petri-dish cultures in which direct observation with the compound 
microscope is feasible. Incubation at 22° to 25° C. will permit the develop- 
ment of satisfactory colonies for descriptive study, although the optimum 
for certain species is above or below this range as will be noted in connection 
with these descriptions and in the general discussion on the influence of 
temperature on colony growth and development in the genus (p. 45). 
Colony descriptions are based upon 3-week old cultures, except as otherwise 
stated. For comparative culture the authors have followed Dale (1909) 
in using substrata containing high concentrations of sugar. The following 
formula is recommended : 

Czapek's solution agar with 20 percent of sucrose 

Sodium nitrate 3 gm. 

Dibasic potassium phosphate 1 gm. 

Magnesium sulfate 0.5 gm. 

Potassium chloride 0.5 gm. 

Ferrous sulfate 0.01 gm. 

Sucrose 200 gm. 

Agar 15.0 gm. 

Dist. water 1,000 cc. 



102 A MANUAL OF THE ASPERGILLI 

Group Limits and Relationships 

Exceptional strains lacking perithecia but presenting conidial morphology 
clearly belonging to the A. glaucus group in its strictest sense are occa- 
sionally found. In addition, the A. reslrictus series (A. penicilloides series 
of George Smith, 1931) shows conidial morphology clearly related to the 
group, but differing markedly from the usual types in colony coloration and 
in the absence of perithecia. Thorn and Church (1926) considered these 
latter types as representing intermediate forms between the Aspergillus 
glaucus and A. fumigatus groups. In the present treatment, we include 
them as non-ascosporic and for the most part diminutive forms sufficiently 
related to the ascosporic species to be included with them in the A . glaucus 
group. 

Certain other groups of the Aspergilli present characters suggesting those 
of the "glaucus" group considered here. The ascospores of A. fischeri 
Wehmer (See A. fumigatus group) and of the A. nidulans group (which 
see; also Thorn and Raper, 1939) in general resemble those of the A. 
glaucus group, but the yellow perithecia suspended by yellow and red 
encrusted hyphae do not occur outside of this group. 

Group Key (Based Primarily Upon Perithecia and Ascospores) 

I. Perithecia present. 

A. Ascospores lenticular, 6m or less in long axis. 

1. Ascospores with convex faces smooth (or nearly so). 

a. Equatorial ridges lacking, furrow absent or showing only as a trace 

A. repens series 

b. Equatorial ridges low and rounded, furrow broad and shallow 

A . ruber series 

c. Equatorial ridges thin and flexuous, crestlike (spore resembling 

a pulley) A. chevalieri series 

2. Ascospores with convex faces rough A. amstelodami series 

B. Ascospores lenticular, 6m or more in long axis 

Large-spored species or the (E.) herbariorum series 

II. Perithecia absent. 

A. Colonies predominantly in yellow-orange to brownish shades. 

1. Heads approximating those of A. repens A. argillaceus Biourge 2 

2. Heads proliferating, giving rise to many subheads 

A. proliferans G. Smith 

B. Colonies in dark green or blue-green shades. 

1. Restricted, velvety, with short conidiophores and abundant columnar 

heads A . reslrictus series 

2. Spreading, floccose, with long conidiophores and globose heads 

A. itaconicus Kinoshita 

2 Culture distributed by Biourge as a new species. Believed to represent only a 
non-ascosporic strain of A. repens, hence not recognized as a valid species by the 
writers (see p. 111). 



THE ASPERGILLUS GLAUCUS GROUP 103 

THE ASPERGILLUS REPENS SERIES 

Ascospores lenticular, mostly 4.8 to 5.4/x by 3.8 to 4.4/x, smooth-walled, 
with equatorial area rounded or somewhat flattened and occasionally in- 
dented showing a trace of furrow, but without crests or ridges. 

The Aspergillus repens series as based upon the ascospore described 
includes a great number of universally distributed strains which retain 
some cultural individuality. Consequently several of them have been 
described as species by earlier workers. From comparison of a great series 
of these forms, it' seems necessary to bring together under the name A . repens 
(Cda.) DeBary, a very considerable number of forms (some of them regarded 
as species by others) in which the ascospore is typical for the group and the 
colony difference falls within lines of quantitative rather than qualitative 
variation. 

The following key is offered as a means of separating culturally distinct 
strains or groups of strains : 

A. Conidial heads large, borne above the surface layer of perithecia and enveloping 

hyphae. 

1. Heads radiate, long-stalked A. repens (Cda.) DeBary 

2. Heads columnar, short-stalked A. dierckxii Biourge 3 

B. Conidial heads small, enmeshed with the perithecia in a felt of sterile hyphae. 

1. Felt orange-yellow, loose-textured, radially wrinkled 

A. pseudoglaucus Bloch. 

2. Felt yellow-buff, close-textured, plane or nearly so A. profusus Hann 3 

Aspergillus repens (Cda.) DeBary, in Abhandl I. Senkenberg. Natiirf. 

Gesellsch. 7:379. 1870. 

Synonyms: A. glaucus var. repens Cda., Icones Fungorum 5: 53, Taf* 

II, fig. 27. 1842. 
A. scheelei Bain, and Sart., Soc. Mycol. de France, Bui. 

Trimest. 28: 257-262, pi. X. 1912. 
A. B var. scheelei Bain, and Sart., Soc. Mycol. de France, 

Bui. Trimest. 28 : 262-267, pi. XI. 1912. 

Colonies upon Czapek's solution agar (3 percent sucrose) restricted, 
plane or somewhat wrinkled, forming a rather compact felt (fig. 24 Ai), 
with the marginal area near Scheele's green (Ridgway, PI. VI) from de- 
veloping heads, older areas yellow-green to greenish-gray and enmeshing 
large numbers of aborted perithecia producing few ascospores; normal 
perithecia found only when such colonies spread over the bare walls of the 
vessel. Reverse in shades of greenish-yellow at colony margin to deep 
maroon or almost black in older areas. 

3 Species name not recognized as valid by authors of this publication. 



104 



A MANUAL OF THE ASPERGILLI 




Fig. 24. Comparative growth of Aspergillus repens, A. chevalieri, and A. ruber 
upon two different culture media; three weeks incubation at room temperature. A, 
1 and#,A. repens, NRRL No. 17, upon (1) Czapek's solution agar (3 per cent sucrose)' 
and (2) Czapek's solution agar with 20 percent sucrose. B, 1 and 2, A. chevalieri 
NRRL No. 78; and C, 1 and 2, A . ruber., NRRL No. 52, upon the same media in similar 
arrangement. 



THE ASPERGILLUS GLAUCUS GROUP 



105 



Colonies upon Czapek's solution agar with 20 percent of sucrose spread- 
ing broadly and rapidly, plane or slightly wrinkled, orange-yellow, com- 
monly characterized by broad zones of dull-green conidial heads (PI. Ill C, 
and fig. 25, A and B) ; surface growth consisting of loosely woven hyphae 
studded with orange granules enmeshing abundant yellow perithecia above 




Fig. 25. Comparative growth of members of the Aspergillus repens series upon 
Czapek's solution agar containing 20 percent of sucrose; incubation at room tempera- 
ture for three weeks. A and B, Typical cultures of A. repens. C, A. pseudoglaucus, 
NRRL No. 40. D, A. pseudoglaucus, NRRL No. 45. 

which project abundant conidial heads, the whole colony and especially 
the marginal areas and adjacent wall of the culture dish commonly over- 
grown by a loose aerial network of hyphae bearing conidial heads and scat- 
tered perithecia; reverse varying from yellow-orange to deep maroon. 

Perithecia very abundant, borne in loose networks of yellow to orange- 
red hyphae (fig. 26 A), yellow, spherical to subspherical, mostly 75 to 100m, 









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Fig. 26. Marginal areas of colonies of representative species of the Aspergillus 
glaucus group showing the relative size and arrangement of perithecia (conidial 
heads not in focus): A, Aspergillus repens;B, A. chevalieri; C, A. ruber; D, A. am- 
stelodatnt; E, A. echinulatus . Figures X 10; inserts X 30. (Reprinted from Thorn 
andRaper, "The Aspergillus glaucus Group," U.S. D. A. Misc. Pub. 426: 1-46. 1941.) 



106 



THE ASPERGILLUS GLAUCUS GROUP 107 

occasionally up to 125/x; asci 10 to 12ju; ascospores lenticular, mostly 4.8 
to 5.6/x by 3.8 to 4.4ju, smooth-walled, with equatorial area rounded or 
somewhat flattened and occasionally indented showing a trace of furrow 
but without crests or ridges (fig. 27 A). Conidial heads abundant, varying 
in different strains from 125 to 175m in diameter, consisting of diverging 
chains of conidia radiating from a hemispherical vesicular apex of the 
conidiophore (fig. 28 A); conidiophores smooth, mostly colorless, 500 to 
1,000/x in length, broadening at the apex to a vesicular area, about 25 to 
40/i m diameter; sterigmata in one series 7 to 10/z by 3.5 to 4.5/u; conidia 
elliptical to subglobose, spinulose, mostly 5 to 6.5/x. 

Represented by cultures NRRL No. 12, No. 17, and more than a score 
of others included in this study. In this connection it should be noted 
that of 37 cultures examined in the present study that produced ascospores 
characteristic of the A . repens series, 29 produced colonies and microscopic 
details that place them in the species A. repens as described. 

This description is manifestly broad enough to include strains approxi- 
mating the description given by Bainier and Sartory for Aspergillus scheelei 
and Aspergillus B var. scheelei (1912b). Evidently A . scheelei was thought 
by the describers to represent a species with somewhat larger ascospores 
showing a more definite furrow, whereas Aspergillus B var. scheelei was a 
strain with smaller ascospores almost without a trace of furrow. Both 
species were described as characterized by the production of a yellow pig- 
ment. In the authors' experience, a distinction based upon color is largely 
invalidated by variants bridging the whole range from yellow-orange to 
deep orange-red and even shades of brown when large numbers of strains 
of this series are compared in culture. Strains also vary slightly in the 
pattern of their ascospores, some rarely producing spores with a trace of 
furrow and others bearing a large proportion with such traces. But among 
spores of a single strain limited variation in this character is normally 
encountered. Thus the presence or absence of a slight furrow, unless 
accompanied by significant differences in morphology or colony character, 
would not seem to justify specific descriptions in this series. 

A strain designated as Aspergillus dierckxii, presumably by Biourge 
but thus far unpublished, was included in Gould and Raistrick's study of 
pigment production in the A. glaucus group (1934). As received from 
Raistrick's laboratory, this organism (NRRL No. 39) produces colonies 
showing no zonate arrangement of conidial heads. Further, heads are 
borne on shorter conidiophores than in typical A. repens and consist of 
columns of conidia rather than radiating chains. Little or no red color 
appears in the colonies or in reverse. Although no other strains showing 
exactly these differences have appeared in the authors' collection, separa- 
tion as a distinct species is believed unwarranted. 



108 



A MANUAL OF THE ASPERGILLI 










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Fig. 27. Ascospores representative of the four small-spored series of the .Aspe? - - 
</i7Jus glaucus group. A, A. repens. B, A. ruber. C, A. chevalieri. D, A. amstelodami. 
In each species upper left and right and center left spores represent surface, profile 
views; center right, surface in face view; lower left, optical section in profile; and 
lower right, optical section in face view. (Reprinted from Thom and Raper, "The 
Aspergillus glaucus Group," U. S. D. A. Misc. Pub. 426: 1-46. 1941.) 






JSSt&- 















xttS?" x?' Conidial structures in the Aspergillus glaucus group, X 750: 4. A. revens, 
NRRL No. 21; 5, A. echinulatus, NRRL No. 131; C, A. restrictus, NRRL No. 154 
ZJ, A. ttacomcus, NRRL No. 161. 

109 



110 A MANUAL OF THE ASPERGILLI 

Aspergillus pseudoglaucus Blochwitz, in Ann. Mycol. 27: 207. 1929; 
emend Thorn and Raper, U.S.D.A. Misc. Publ. No. 426, p. 12. 1941. 

Colonies upon Czapek's solution agar (3 percent sucrose) restricted in 
growth, radiately wrinkled, yellow-green to shades of gray, consisting of a 
mixture of small conidial heads, young or aborted perithecia and more or 
less colorless hyphae ; reverse orange at center becoming lighter toward the 
margin. 

Colonies upon Czapek's solution agar with 20 percent of sucrose spread- 
ing, strongly wrinkled in a predominantly radiate manner, consisting of a 
felt of orange-encrusted hyphae enmeshing abundant perithecia, orange 
except at margin where yellow-green predominates from the presence of 
small conidial heads admixed with perithecia in the mycelial felt (fig. 25 C) ; 
reverse yellow becoming orange-brown or maroon in marginal areas. 

Perithecia abundant, spherical to subspherical, mostly 60 to 80m though 
occasionally 100m in diameter, yellow, embedded in a felt of orange my- 
celium; asci 10 to 12/i in diameter; ascospores lenticular, 4.6 to 5.2/x by 
3.6 to 4.0m, occasionally 5.6m m long axis, smooth-walled, with equatorial 
region rounded or flattened, without ridges, and with furrow generally 
lacking though occasionally showing as a trace. Conidial heads few in 
number and generally submerged in the mycelial felt, small, mostly 50 to 
75m in diameter but occasionally up to 100m; conidiophores mostly 150 to 
300m in length, 5 to 8m at the base, broadening to a terminal vesicle 12 to 
20m in diameter; sterigmata in a single series, 6 to 8m by 3 to 4m; conidia 
subglobose, delicately spinulose, variable in size ranging from 5.5 to 7.5m 
in diameter. 

Represented in the NRRL collection by No. 40 received from Baarn 
as A. pseudoglaucus Blochwitz and No. 41 received from George Smith as 
A . fumigatoides Bain, and Sart. 

There is reason to believe that the former culture is directly derived from 
Blochwitz 's type. It becomes necessary therefore to emend the description 
given by him insofar as the measurements and markings of ascospores and 
conidia are concerned. Gould and Raistrick (1934) reported biochemical 
data upon a culture, No. A 38, received from Biourge as A. fumigatoides 
Bain, and Sart. (NRRL No. 41), which is identical with A. pseudoglaucus 
(NRRL No. 40) as sent to the authors by Westerdijk. Obviously the cul- 
ture from Biourge is incorrectly named. It does not fit the species descrip- 
tion nor the figures of A. fumigatoides (1909) in the size of its conidia, the 
character of its perithecial wall, or the pattern of its ascospores. The asco- 
spores of A . fumigatoides are shown as roughened over their entire surfaces, 
as in A. fischeri, whereas those of No. 41 certainly belong in the A. repens 
series. Distinctive colony characters, however, maintained stably through 
many transfers, together with the size of the conidial apparatus, warrant 
separating A. pseudoglaucus from A. repens and maintaining it as a species. 



THE ASPERGILLUS GLAUCUS GROUP 111 

A strain, NRRL No. 45, received from Dr. B. O. Dodge and Miss Mar- 
jorie E. Swift, of the New York Botanical Garden, in 1931, is characterized 
by an intensely wrinkled colony and a further reduction in the size and 
number of conidial heads (fig. 25 D). Colonies are dull orange red and bear 
abundant perithecia enmeshed in a close felt of sterile encrusted hyphae. 
Obviously it should be considered with A. pseudoglaucus. 

In cultures received from Baarn (NRRL No. 44) and from George Smith 
(NRRL No. 45) as A . profusus Hann (nomen nudum) there is a pronounced 
accentuation of the floccose habit already noted in A. pseudoglaucus. 
Upon 20 percent sucrose Czapek agar these cultures, which are obviously 
duplicates, produce spreading, plane or radiately wrinkled, floccose colonies 
consisting of a close felt of light tan to buff -colored hyphae, bearing occa- 
sional perithecia and widely scattered conidial heads. The perithecia are 
commonly embedded deep within the felt, whereas the conidial heads are 
most evident at the colony margin. Although the ascospores of these 
strains are definitely of the A. repens type, they are generally flattened 
along their equators and commonly show a trace of furrow. An occasional 
ascospore shows a minute roughness in the equatorial region. The dif- 
ferences observed do not seem to warrant perpetuating the name A. pro- 
fusus, and in agreement with Dr. Westerdijk and coworkers (Centraal- 
bureau List, 1939), the cultures have been assigned to A. pseudoglaucus. 

Culture NRRL No. 46, received from Raistrick in 1923 as Aspergillus 
novus Wehmer (nomen nudum) bears ascospores duplicating those of the 
cultures just considered. This strain is of particular interest, because in 
routine transfers colonies of two distinct types commonly appear. One 
of these is predominantly floccose and suggests the colonies of the strains 
received as A . profusus. The other consists of a crowded surface layer of 
perithecia, which is thinly veiled by a loose felt of orange-red hyphae, and 
in its gross appearance, with the exception of its lighter color, is strongly 
suggestive of certain cultures of Aspergillus ruber. The authors agree with 
Wehmer (1901) and Blochwitz (1929a) that the species designation, Asper- 
gillus novus, should be withdrawn. 

A nonascosporic culture distributed by Biourge as Aspergillus argillaceus 
n. sp., was received upon two occasions from Prof. Raistrick's laboratory. 
From its appearance in culture and the morphology of its conidial struc- 
tures this fungus would seem to represent a member of the Aspergillus 
repens series in which perithecial development has been wholly suppressed. 
Although it is questionable whether this fungus represents a true species, 
a brief description is given because of its inclusion in biochemical studies 
by Raistrick and coworkers (1939, 1934, 1937). Colonies upon Czapek's 
solution agar with 20 percent of sucrose spreading irregularly, consisting of 
a loose floccose felt of aerial hyphae and abundant conidial heads, pale 
yellow-green to clay color; reverse yellow. Upon Czapek's solution agar 



112 A MANUAL OF THE ASPERGILLI 

(3-percent sucrose) colonies restricted, raised in center, thinning toward 
margin, consisting of abundant conidial heads and interlacing hyphae, buff 
to clay colored; reverse yellow to tawny. Conidial heads abundant, dull 
green, up to 200m in diameter and commonly splitting into fairly well-de- 
fined columns, conidiophores up to 1,000m in length. Conidia subglobose 
mostly 5.5 to 6.0m but occasionally up to 7.0m in long axis, spinulose. 

ASPERGILLUS RUBER SERIES 

Ascospores lenticular, 5.0 to 6.0m by 4.0 by 4.8m, colorless, with broad, 
shallow furrow generally evident and flanked by low ridges, and with walls 
smooth except for minute roughness along the equatorial ridges. 

This series includes a great number of strains showing variations in cul- 
tural appearance but producing ascospores of a limited size range and 
fairly well-defined pattern. For this particular study some 30 strains, 
received from various culture collections and contributors and selected 
from the isolations made in this laboratory over a period of many years, 
have been chosen for repeated culture and examination. Among these, 
many strains appear distinct, but their differences are commonly bridged by 
intermediate forms. Separation within the series, therefore, must be along 
one of the following lines — either (1) strains must be separated upon minor 
characters, such as differences in the intensity of pigmentation, slight 
variations in ascospore character, etc.; or (2) strains must be set off in 
broad and elastic subgroups, in some cases including large numbers which 
vary appreciably in detail. The second alternative is desirable, for the 
first can lead only to increased hairspliting and end in greater confusion 
than that which already exists. 

Spieckermann and Bremer's designation Aspergillus ruber (1902) is as- 
signed to the series, because its members are predominantly producers of 
an intense red pigment and bear ascospores of the general size and pattern 
described by these authors. The only other described fungus possessing a 
similar ascospore and characterized by its red color is Bainier and Sartory's 
Aspergillus sejunctus (1911b). 

Although it is now quite impossible to say what particular fungus either 
pair of investigators had at hand, both are believed to have worked with 
members of the large series now under consideration. Aspergillus ruber 
is retained since its description is more adequate for the series in addition 
to being the prior species. 

To accentuate cultural similarities and differences between the strains 
studied, a wide variety of culture metiia has been employed. Based upon 
their appearance in culture, the strains fall into a few well-defined sub- 
groups (see figs. 29 and 30). The strains belonging to one of these seem 



THE ASPERGILLUS GLAUCUS GROUP 



113 



to represent the fungus described by Spieckermann and Bremer (1902) and 
are at the same time most abundant in the entire series, hence are consid- 
ered as typical of A. ruber (figs. 29 A and 30 A). The general characters of 
the remaining subgroups are listed to show the extreme cultural variation 




Fig. 29. Different colony types developed in the Aspergillus ruber series in three 
weeks at room temperature upon 20 percent sucrose Czapek agar : A , Typical A . ruber, 
NRRL No. 52; B, NRRL No. 70, characterized by thin colonies, with mycelium 
largely submerged; C, NRRL No. 65, colony floccose, bearing abundant perithecia 
and few conidial heads; andZ), NRRL No. 75, deep floccose colony with very abundant 
conidial heads and only scattered perithecia. 



that is to be expected within the series, but specific names are withheld, 
because to perpetuate or propose such would multiply rather than clarify 
the confused nomenclature of this abundant and variable series of organ- 
isms. 



114 A MANUAL OF THE ASPERGILLI 

To illustrate more definitely the variation that occurs, the following 
outline of possible lines of separation is inserted: 

A. Colonies predominantly perithecial. 

1. Colonies red, perithecia abundant in a layer at the agar surface and over- 

grown by a felt of red encrusted hyphae 

A. ruber (Spieck. and Brem.) Thorn and Church, NRRL Nos. 52, 49 

2. Perithecia abundant in a loose, floccose overgrowth of red hyphae as well 

as in a layer at the agar surface NRRL No. 65 

3. Colonies thin, orange-red, perithecia abundant in old cultures and on very 

concentrated media NRRL No. 70 

B. Colonies predominantly conidial. 

1. Colonies gray, heads long-stalked, perithecia few NRRL No. 75 

C. Colonies mixed conidial and perithecial. 

1. Colonies orange-red and green, zonate, perithecia borne at the agar surface 
and piled in a loose network of superficial hyphae NRRL No. 71 

D. Colonies predominantly floccose, colonies red-brown, perithecia and conidial 

heads few ^4. lovaincnsis Biourge, 4 NRRL No. 76 

Aspergillus ruber (Bremer) 

Synonyms: A. ruber (Spieckermann and Bremer) Thorn and Church, in 

The Aspergilli, 112. 1926. 
Eurotium rubrum Bremer, in Zeitschr. f. Untersuch. d. Nah- 

rung. und Genussmittel IV. 1901, p. 72, also in Die fett- 

verzehr. Organismen in Nahr. u. Futtermitteln, Dissert. 

Munster 1902. 
E. rubrum Spieckermann and Bremer, in Landw. Jahrb. 31 : 

81-128. 1902. 
A. sejunctus Bain, and Sart., Soc. Mycol. de France, Bui. 

Trimest. 27: 361-367, pi. XL 1911. 

Colonies upon Czapek's solution agar (3 percent sucrose) more restricted, 
plane (fig. 24 Ci), orange-brown to red-brown in color; perithecia generally 
abundant though often abortive; conidial heads pea-green to olive-green, 
abundant in some strains, few and largely vestigial in others ; reverse orange- 
red to maroon. 

Colonies upon Czapek's solution agar with 20 percent of sucrose spread- 
ing rapidly and broadly in a regular manner or unevenly, plane, predom- 
inantly red, ranging from ferrugineous to morocco red; perithecia very 
abundant, borne in a dense layer at the agar surface and largely concealed 
within and beneath a close-textured felt of red-encrusted hyphae; conidial 
heads projecting above the felt, pale gray-green to deep olive-gray, more or 
less abundant, and generally crowded near the center or scattered unevenly 

4 Species name not recognized as valid by authors of this publication. 



THE ASPERGILLUS GLAUCUS GROUP 



115 



over the colony (PI. Ill D; figs. 29 A and 30 A); reverse in shades of dark 
red-brown. 



**m3& .& m® WW #M M 



_cor» 




Fig. 30. Diagramatic representation of cross sections of different colony types 
in the Aspergillus ruber series developed at room temperature upon 20-percent sucrose 
Czapek agar, showing relative abundance and disposition of the conidial heads (con) 
and perithecia (per), and the amount and character of the mycelium (my) above the 
substratum (sub) : A, Typical colony of A. ruber as seen in NRRL No. 52; B-F,atypica 
colonies as seen respectively in NRRL No. 71, No. 70, No. 65, No. 75, and No. 76. 
Scale approximate. (Reprinted from Thorn and Raper, "The Aspergillus glaucus 
Group," U.S.D.A. Misc. Pub. 426: 1-46. 1941.) 

Perithecia very abundant, largely enmeshed in a felt at the agar surface 
(fig. 30 A), yellow to orange-red, spherical to subspherical, mostly 80 to 
120m though occasionally up to 140m in diameter; asci 12 to 15m; asco- 
spores lenticular, 5.2 to 6.0m by 4.4 to 4.8m, with furrow generally evident 
as a broad and shallow depression around the spore equator, ridges low and 



116 A MANUAL OF THE ASPERGILLI 

often inconspicuous, walls smooth except for minute roughness along 
equatorial ridges (fig. 27 B). Conidial heads generally abundant, numerous 
in localized areas or scattered thinly over the colony, pale blue-green, 
radiate, 150 to 250m in diameter; conidiophore smooth, colorless to orange- 
brown, 500 to 750m in length, broadening to 14 to 16m where it passes into 
the subglobose vesicular area of 25 to 35m diameter; sterigmata in a single 
series 7 to 9m by 4 to 5m; conidia elliptical to subglobose, closely spinulose, 
mostly 5 to 6.5m in long axis. 

Aspergillus ruber is represented in this study by NRRL Nos. 52, 53, and 
others. Of 31 strains examined belonging to the whole series, 19 showed 
colonies and microscopic characters that place them within the species 
Aspergillus ruber as described above. Although the majority of strains 
belonging to the A . ruber series produce plane colonies as noted in the de- 
scription, occasionally strains may produce colonies more or less wrinkled. 

Culture NRRL No. 65 (figs. 29 C and 30 D) represents a subseries of 
several strains that differ from the above not only in colony character upon 
20 percent sucrose Czapek agar, as indicated in the preceding key, but also 
in their growth upon media of lower concentration. These grow slowly 
and poorly on Czapek (3 percent sucrose), potato-dextrose, and wort 
agars, producing small raised colonies of 1 to 2 cm. in diameter bearing 
neither normal conidial heads nor perithecia. 

Strain NRRL No. 70 (figs. 29 B and 30 C) produces abundant perithecia 
only on very dry areas of the substratum in old cultures or on media con- 
taining a sucrose concentration of 40 percent or more. In contrast to other- 
strains, this fungus grows better upon media containing 4 percent agar than 
the usual 1.2 percent agar, further establishing its xerophytic character. 

Strains such as NRRL No. 75 (figs. 29 D and 30 E), occasionally en- 
countered, are predominantly conidial and characterized by rampant 
hyphae bearing abundant conidial heads piled in floccose masses above the 
substratum and upon the edges of the culture dish or tube. They thus 
produce colonies markedly in contrast with the usual Aspergillus ruber 
concept. But the character of their ascospores, together with the occur- 
rence of occasional sectors in colonies of these strains showing the usual 
mixture of perithecia and conidial heads, relates them definitely with A. 
ruber. 

Strain NRRL No. 71 (fig. 30 B) represents a subsection of the series 
in which conidial heads are abundant and generally arranged in fairly 
definite zones and patches with loose clusters of perithecia irregularly and 
conspicuously distributed among and above the grouped green heads. 
These strains are further characterized by somewhat larger perithecia than 
those of NRRL No. 52, being mostly in the range of 125 to 150m in diameter, 
and by producing less red color in the colonies and in their reverse. 



THE ASPERGILLUS GLAUCUS GROUP 117 

Culture NRRL No. 76 (fig. 30 F) is characterized by a close felt of 
red-brown hyphae, which completely covers the agar surface and in 
which scattered perithecia are borne. Conidial heads are scarce and 
largely confined to the colony margin. This strain, which was included in 
Gould and Raistrick's study of pigmentation in the Aspergillus glaucus 
group (1934), was received from George Smith under the name Aspergillus 
lovainensis and attributed to Biourge. Except for its dark color this 
fungus in culture bears a striking resemblance to one received from Baarn 
as Aspergillus profusus (NRRL No. 44), which showed similar floccose 
habits. However, the ascospores of the latter are smaller and less furrowed 
and are essentially smooth along the equatorial margin. The degree of 
relationship between the two is questionable. 

In addition to the ascosporic strains definitely placeable in the series, 
George Smith has recently described A. proliferans in which the prolifera- 
tion of the sterigmata has become so pronounced as to become the most 
conspicuous character, while perithecium formation has been suppressed. 
In colony characters, however, it belongs here. This strain diverges 
further from the type as described but may be arbitrarily placed here by 
the branching of its simplified heads and the size and markings of its 
conidia. 

Aspergillus proliferans George Smith, in Brit. Mycol. Soc. Trans. 26(1/2): 

26, PI. III. 1943. 

Colonies on Czapek's solution agar spreading very slowly, with growth 
at first largely submerged, then with matted floccose aerial mycelium, white 
changing to yellowish shades, sporing tardily, with conidial areas gray- 
green, reverse yellowish-brown; on wort agar growing slowly but better 
than on Czapek, with mycelium white then yellow and finally orange and 
tardy development of gray-green to gray conidial areas, becoming more 
deeply floccose in age especially at shallow end of the slope ; reverse yellow ; 
normal conidial heads loosely radiate; conidiophore smooth, thin-walled, 
usually with one or two septa, 4 to 14m in diameter; vesicles occasionally 
almost globose, more frequently obconical or mere broadening of the ends 
of the conidiophores, up to about 20m in diameter; sterigmata when normal, 
in one series, 8 to 1 lju. by 3.5 to 6m, often elongate, septate and bearing 
small secondary heads, frequently resembling heads of monoverticillate 
Penicillia, or with upper portion much swollen and appearing almost as 
very large, thick-walled conidia with long connectives, up to 20m in di- 
ameter, with normal and swollen sterigmata often appearing in the same 
head; conidia globose or subglobose, rough, fairly dark-colored, 5 to 9.5m 
in diameter; perithecia not found. (Species description after George 
Smith.) 



118 A MANUAL OF THE ASPERGILLI 

Aspergillus halophilus Sartory, Sartory, and Meyer (in Ann. Mycol. 28(3/4): 362- 
363, PI. III. 1930) appears from the description based upon colonies grown upon 
licorice sticks, to have been some member of this general group. No cultures have 
been available for comparison, hence placement near A. proliferous of George Smith 
can be only tentative. If placement were to be based upon their figure 12, it might 
be a species of Scopulariopsis. 

ASPERGILLUS CHEVALIERI SERIES 

Ascospores lenticular, mostly 4.6 to 5.0> by 3.4 to 3.8m, occasionally up 
to 5.2ju in long axis, with walls smooth or slightly rough, with crests prom- 
inent, flexuous, often recurved, and with furrow conspicuous but consisting 
more of a trough between extended equatorial crests than a depression in 
the spore wall. 

Strains belonging to this series show appreciable difference in colony 
character and to a limited degree in the surface markings of their asco- 
spores. The ascospores of all, however, are characterized by their con- 
tinuous, prominent equatorial crests which do not form an integral part 
of the spore wall, but extend well beyond the margin of the spore body 
proper. To use Mangin's exceedingly descriptive term, they are charac- 
teristically "pulley-form." 

The following key will serve to differentiate groups of strains within the 
series : 

A. Ascospore walls smooth. 

1. Crests prominent, thin, flexuous, often recurved 

A. chevalieri (Mangin) Thom and Church 

2. Crests evident, low, usually erect 

A. chevalieri var. rnultiascosporus Nakazawa et al 5 

B. Ascospore walls more or less roughened. 

1. Crests thin, flexuous, often recurved; conidia roughened. A. oriolus Biourge 5 

2. Crests thicker, usually erect: conidia smooth 

A. chevalieri var. intermedins, Thom and Raper 

Aspergillus chevalieri (Mangin) Thom and Church, The Aspergilli, p. 11. 

1926. 

Synonym: Eurotium chevalieri Mangin, Ann. des Sci. Nat., Bot. (Ser. 9) 
10:361-362, fig. 12. 1909. 

Colonies upon Czapek's solution agar (3 percent sucrose) restricted, 
plane, closely felted, bluish-gray in center, with typical heads and peri- 
thecia largely confined to marginal area (fig. 24 Ci); reverse maroon in 
center to orange at margin. 

Colonies upon Czapek's solution agar with 20 percent of sucrose growing 
best at 30° C. or above, spreading, plane to somewhat wrinkled in central 

5 Species name not recognized as valid by authors of this publication. 



THE ASPERGILLUS GLAUCUS GROUP 



119 



area (fig. 31 A) with abundant conidial heads in bli'e-green shades, dis- 
tributed evenly over the whole surface or more crowded in localized areas, 
projecting above a layer of abundant perithecia enmeshed in orange-red 
hyphae at the agar surface; reverse in shades of orange-red to brown, more 
intense in center. 

Perithecia abundant and closely enmeshed in a felt of orange-red en- 
crusted hyphae (fig. 26 B), mostly 100 to 140m, occasionally up to 150^, 
globose to subglobose, yellow to orange; asci 9 to 10/z; ascospores lenticular, 
4.6 to 5.0/x by 3.4 to 3.8/z, with walls smooth, with equatorial crests prom- 
inent, thin and often recurved, and with furrow consisting more of a trough 
between parallel crests than an equatorial depression in the spore body 




Fig. 31. Comparative growth of members of the Aspergillus chevalieri series in 
three weeks at room temperature upon 20 percent sucrose Czapek agar: A, Typical 
A. chevalieri, NRRL No. 78; B, A. chevalieri var. intermedins, XRRL No. 82. 



(fig. 27 C). Conidial heads abundant, pale blue-green, appearing radiate 
from divergent conidial chains, mostly 125 to 175^ in diameter, occasion- 
ally larger; conidiophores mostly 700 to 850m in length, enlarging to a 
vesicular apex, somewhat globose, 25 to 35/z in diameter; sterigmata in a 
single series, closely packed, 5 to 7/x by 3 to 3.5/x; conidia subglobose, 
spinulose, mostly 4.5 to 5.5m in diameter. 

Aspergillus chevalieri is represented in the present study by cultures 
XRRL Xos. 78, 79, and others. The species name is limited to strains 
bearing ascospores with smooth walls and prominent, thin equatorial crests 
because it is believed that these strains most nearly represent the organism 
described by Mangin (1909). When grown upon 20 percent sucrose Czapek 
agar, these strains are further characterized by their predominantly orange- 



120 A MANUAL OF THE ASPERGILLI 

red colonies, pale blue-green conidial heads, and dark-colored reverse. 
Within the series, different strains vary in the quantity of conidial heads 
produced, e.g., NRRL No. 79 regularly produces an abundance of heads, 
NRRL No. 78 relatively few. 

Strains of this series are not so commonly encountered as are those of 
the A. repens, A. amstelodami, or A. ruber series. And within the series, 
strains that conform with the typical species description are relatively less 
numerous than in these other series. In the present study 14 strains be- 
longing to the A. chevalieri series have been examined and only 6, or less 
than half, are wholly representative of the species A. chevalieri. 

The series as a whole seems to be relatively unstable, and certain strains 
and groups of strains appear transitional between this series and the ^4. 
repens series on the one hand and the A. amstelodami series on the other. 

Culture NRRL No. 88 received from Baarn as the type of Aspergillus 
chevalieri var. multiascosporus Nakazawa, Takeda, Okada, and Simo points 
toward the A. repens series. Its ascospores have smooth walls, as in the 
typical strains of A. chevalieri, but bear low, erect crests in constrast to 
the thin, flexuous crests characteristic of this species. Spores lacking crests 
are occasionally seen and these closely resemble A. repens. The colony 
upon 20 percent sucrose Czapek agar is definitely of the character of A. 
chevalieri. Nakazawa and coworkers (1934) separated it from A. chevalieri 
because of its more floccose habit and its more abundant production of 
perithecia. The former character is evident in the authors' cultures, but 
perithecia are not produced more abundantly than in certain strains en- 
tirely typical of A. chevalieri. 

Another variation from the typical species is seen in culture, NRRL 
No. 87 received from George Smith as Aspergillus oriolus and attributed 
to Biourge. The ascospores of this culture (and another that is in the 
NRRL collection, No. 81) have crests typical of A. chevalieri, but the spore 
walls are finely roughened over their entire surfaces. This character is 
suggestive of A. amstelodami, although the roughening of the wall is slight 
in comparison with that species. The colony upon 20 percent sucrose 
Czapek agar is essentially like that of typical strains of A. chevalieri but is 
less red in color and bears fewer conidial heads. Although these cultures 
can be distinguished from type they are not recognized as warranting sep- 
aration. 

The roughening of the ascospore wall is further accentuated in a group 
of four apparently similar strains, which it is believed are truly inter- 
mediate between A. chevalieri and A. amstelodami. Because their asco- 
spores bear crests of the A. chevalieri type and hence appear "pulley-form" 
they are retained in the species. However, because they differ from typical 
strains in additional particulars, they are considered a new variety, namely 
Aspergillus chevalieri var. intermedium. 



THE ASPERGILLUS GLAUCUS GROUP 121 

Aspergillus chevalieri (Mangin) var. intermedins Thorn and Raper, in U. S. 
D. A. Misc. Publ. No. 426, p. 21. 1941. 

Colonies upon Czapek's solution agar with 20 percent sucrose differing 
from the species in texture and color, and presenting withal a picture 
intermediate between A. chevalieri and .4. amstelodami (fig. 31 B). Asco- 
spores lenticular, mostly 4.6 to 5.2m by 3.6 to 4.0m, occasionally 5.4m in 
long axis, with walls roughened and with prominent equatorial crests. 
Conidiai heads dull green, radiate to columnar, mostly 100 to 125m in 
diameter, and up to 175m in length ; conidia elliptical to subglobose, smooth- 
walled, mostly 3 to 4m in long axis. 

Represented in this study by culture NRRL Xo. 82 which was received 
from George Smith as No. 107 and bore the following notation: "Isolated 
G. S. from cotton yarn, 1927. Close to .4. chevalieri— differs in having 
smooth, small conidia and ascospores somewhat larger than type." Dupli- 
cated by three additional strains received from European sources. 

Aspergillus chevalieri var. intermedins appears to be transitional between 
the A. chevalieri and the A. amstelodami series. Such a view is supported 
(1) by the pattern of the ascospores, which shows both the extended and 
often recurved equatorial crests characteristic of .4. chevalieri and the 
rough spore walls of A. amstelodami; and (2) by the coloration of the 
colony. Aspergillus chevalieri var. intermedins upon 20 percent sucrose 
Czapek agar becomes orange-yellow above and orange to light brown in 
reverse, A. amstelodami remains bright yellow with reverse uncolored, 
whereas A. chevalieri becomes red in the colony and reverse. The smooth- 
ness of conidia in A. chevalieri var. intermedins is a distinctive character and 
appears in neither A . chevalieri nor A . amstelodami. Although this variety 
from many points of view appears to be a hybrid, proof of such origin is 
lacking. 

Aspergillus diplocystis (Sartory, Sartory, Hufschmitt and Meyer) Dodge, Med. 
Myc. p. 625. 1935. Syn. Eurotium diplocyste Sartory, Sartory, Hufschmitt and 
Meyer, in Compt. Rend. Soc. Biol. 104: 881-883. 1930. Not E. diplocystis B. and 
Br., Jour. Linn. Soc. 14: 55-56 Tab. 10. 1875. 

Characterization: Colonies greenish-yellow, becoming yellow from perithecia. 
Conidiophores erect , 50 to 100m high, 3.1 to 3.7m in diameter, membrane thick, hyaline. 
Sterigmata confined to a portion of head, 5 to 6.25m by 1.5 to 2.5m- Secondary sterig- 
mata small ; conidia spherical, 2.25 to 3.1m in diameter, slightly ellipsoid, green (tendre 
to cendre); sterigmata sometimes abortive and proliferous. Perithecia canary 
yellow asci 4 to 6m by 5 to 7m, containing 8 ascospores which are ovoid, with a furrow 
and two crests, 1.5 to 2.5m by 1.8 to 3.1m- 

This description suggests an Aspergillus with the heads approximating .4 . nidulans 
and the perithecia of A. chevalieri. Ascospore measurements as reported are appre- 
ciably smaller than those of A. chevalieri or any other known species of Aspergillus. 
It was described from a case of onychomycosis from the thumb and the great toe. 
Tentatively placed in the A. chevalieri series. The name is invalid because of E. 
diplocystis B. and Br. 1S75. 



122 A MANUAL OF THE ASPERGILLI 



ASPERGILLUS AMSTELODAMI SERIES 



Ascospores 4.7 to 5m by 3.6 to 3.8^, lenticular, colorless, with equatorial 
furrow conspicuous, broadly V-shaped and flanked by broad irregular 
ridges, with walls irregularly and unevenly ridged or roughened over the 
entire surface. 

Included in this series are strains that differ greatly in colony appearance. 
However, their close relationship is demonstrated by the similarity in size 
and pattern of their ascospores and is further shown by the dark olive-green 
color of their conidial heads, the bright yellow color of their perithecia, 
and the absence of any red either in the colonies or their reverse. 

The following key is designed to show the variation that occurs within 
the series and to offer a means of separating strains or groups of strains 
that are culturally distinct : 

A. Colonies predominantly perithecial. 

1. Conidial heads abundant in central area and often in concentric zones 

A. amstelodami (Mangin) Thorn and Church 

2. Conidial heads widely scattered or lacking NIHIL No. 113 

B. Colonies predominantly conidial. 

1. Perithecia widely scattered, superficial NRRL No. Ill 

2. Perithecia abundant in a felted layer above the conidial heads 

A.montevidensis Talice and MacKinnon 

C. Colonies very thin, perithecia and conidial heads widely scattered 

NRRL No. 110 

Aspergillus amstelodami (Mangin) Thom and Church, The Aspergilli, p. 

113. 1926. 

Synonyms: Eurotium amstelodami Mangin, in Ann. des Sci. Nat., Bot. 
(ser. 9) 10: 360-361. 1909. 
E. repens var. amstelodami Vuill., Soc. Mycol. de France, 
Bui. Trimest 36: 131. 1920. 

Colonies upon Czapek's solution agar (3 percent sucrose) restricted, 4 
to 6 cm. in diameter, plane or closely wrinkled, yellow to dull yellow-gray 
in color from abundant perithecia admixed with sterile hyphae and de- 
veloping conidial heads; reverse uncolored, becoming tawny in age. 

Colonies upon Czapek's solution agar with 20 percent of sucrose spread- 
ing, 8 to 10 cm. in diameter, more or less wrinkled and zonate (PI. Ill E, 
and fig. 32 A), perithecia very abundant and clustered in masses forming 
a dense layer at the agar surface (fig. 26 D), bright yellow in color, lending 
a characteristic appearance to the colony; conidial heads deep olive-green, 
abundant in colony center and scattered more or less unevenly over the 
whole surface, occasionally obscuring the layer of perithecia beneath. 



THE ASPERGILLUS GLAUCUS GROUP 



123 



Reverse persistent^ yellow under perithecial areas, more or less green 
where conidial areas predominate. 

Perithecia globose to subglobose, mostly 115 to 140/x in diameter, occa- 
sionally up to 160m, not covered by or embedded within a felt of sterile 
hyphae (fig. 26 D); asci mostly 10 to 12/z, 8-spored; ascospores lenticular, 




Fig. 32. Comparative growth of members of the Aspergillus amstelodami series: 
A, Typical A. amstelodami, NRRL No. 90; B, NRRL No. 113, a strain producing 
abundant perithecia and few conidial heads; C, NRRL No. Ill, strain producing 
abundant conidial heads and very few perithecia; D, A . montevidensis , NRRL No. 108. 

4.7 to 5.0m by 3.6 to 3.8/;, with prominent V-shaped equatorial furrow and 
broad irregular ridges, and with walls roughened over their entire surfaces 
(fig. 27 D). Conidial heads radiate-columnar, mostly 120 to 150m in 
diameter, occasionally larger; conidiophores colorless to pale yellow-green, 
275 to 350m in length, broadening to 10 to 12m in diameter below the 
vesicle; vesicle subglobose, 18 to 25m in diameter; sterigmata about 5 to 



124 A MANUAL OF THE ASPERGILLI 

6.5/x by 2.5 to 3.5^; conidia finely spinulose, subglobose, variable in size, 
ranging from 3.5 to 5.2/u mostly about 4/x in long axis. 

Represented in the XRRL collection by cultures Xos. 89, 90, and many 
others. 

Thirty-two cultures belonging to this series have been examined in the 
present study. Included in this number are the authors' own isolates from 
a wide variety of sources together with cultures contributed by collabora- 
tors in this country and abroad. Of these, more than three-fourths regu- 
larly produce colonies conforming with the above description of the species 
A. amstclodami. Although wide variation in colony character does occur 
within the series, it is obvious that such variations are exceptional rather 
than commonplace. Accordingly, it is not believed advisable to assign or 
create specific or varietal names for these variations although they differ 
markedly from the typical A. amstclodami in gross appearance. An excep- 
tion to this policy has been made in the case, of cultures received as A. 
montevidcnsis, for reasons that will be considered later. 

As indicated in the preceding key to the series, marked variation from 
the normal cultural character of .4. amstclodami occurs along certain diver- 
gent lines. 

Culture XRRL Xo. 113 (fig. 32 B) received from Baarn as Eurotium 
repcns (Cda.) DeBary and Wor. var. amstclodami Vuill. (1920) represents 
a variation that tends toward an almost complete suppression of the conidial 
phase with only an occasional small and atypical head present. 

In the opposite direction, culture XRRL Xo. Ill (fig. 32 C) recently 
received from Bliss in California (isolated from date fruits) represents a 
variation that produces a dense stand of conidial heads and only occasional 
perithecia, these being borne above rather than below the layer of crowded 
conidial heads. 

In contrast to both of the preceding, culture XRRL Xo. 110 isolated 
from an old shoe, produces an extremely thin, spreading colony that bears 
only widely scattered perithecia or conidial heads. 

A fourth distinct variation is represented by culture XRRL Xo. 108 
received in 1932 from Talice as A. montevidcnsis Talice and MacKinnon 
(1931). This fungus is characterized by an initially strong development of 
the conidial phase, and subsequently of perithecia in a felted overgrowth, 
which in the colony center more or less obscures the underlying conidial 
layer. Perithecia and conidial heads are somewhat smaller than in strains 
of A. amstclodami. Although this culture does not differ from ^4. amstelo- 
dami more widely than the variations previously noted, since it has an im- 
puted pathogenic history and since it has been described and distributed 
widely under the name Aspergillus montevidcnsis, it is believed advisable 
to retain the name in association with this culture. Accordingly, the 
writers include the following emended description. 



THE ASPERGILLUS GLAUCUS GROUP 125 

Aspergillus montevidensis Talice and MacKinnon, in Soc. de Biol. (Paris) 

Compt. Rend. 108: 1007-1009. 1931. emend. Thorn and Raper, 

U. S. Dept. of Agr. Misc. Pub. 426, p. 20. 1941 . 

Colonies upon Czapek's solution agar (3 percent sucrose) restricted, 
radiate sulcate, with zonation evident toward the margin, central area 
showing coremia, perithecia few or lacking; reverse and agar very dark, 
almost black. 

Colonies on Czapek's solution agar with 20 percent of sucrose, spreading, 
wrinkled and buckled (fig. 32 D), at first bluish-green from massed conidial 
heads, with central area later becoming yellow from developing perithecia 
in a more or less tufted overgrowth of somewhat floccose mycelium ; reverse 
in yellow-green shades to deep olive in colony center. 

Perithecia abundant, of variable size and irregular shape with relatively 
few fertile asci and ascospores, late in developing, commonly 75 to 100m 
in diameter, occasionally larger; asci 10 to 12/x in diameter; ascospores 
lenticular, roughened, with broad and prominent furrow flanked by low 
acute and irregular ridges, mostly 4.8 to 5.2m by 3.6 to 4.0m, occasional 
spores larger or smaller. Conidial heads very abundant, small, somewhat 
columnar, with few conidial chains, mostly 70 to 80m wide, occasionally 
up to 100m; conidiophore up to 300 to 350m long, frequently very short 
when borne upon the aerial mycelium, broadening to a hemispherical dome- 
like vesicular area at the apex; commonly deep green or greenish-brown; 
vesicle mostly 15 to 20m in diameter, occasionally larger or smaller; sterig- 
mata in one series relatively short and thick, 6 to 7m by 3 to 3.5m; conidia 
roughened, subglobose, commonly 4 to 5m by 3 to 4m, occasionally 5.5m 
diameter. 

Type culture isolated by Talice and MacKinnon from the tympanic 
membrane of the human ear (1931). It is carried in the XRRL collection 
as No. 108. 

LARGE-SPORED SPECIES, OR THE HERBARIORUM SERIES 

Under Eurotium herbariorum Lk., Mangin includes all of the members 
of the group with ascospores more than 6.6m in long axis (1909). In a 
general way this represents a very common usage in older literature begin- 
ning as far back as Corda in the 1830's. Because neither measurements 
nor markings of the ascospores were given, no one can fix the type of E. 
herbariorum. In general, the species in the large-spored group have both 
conidia and ascospores definitely larger than those in series already de- 
scribed. They become very conspicuous to the collector who finds the 
anomalous situation of an overabundance of published names and a dearth 
of isolations. Over a period of many years the scarcity of strains isolated 
in this laboratory which show ascospores larger than 7.0m leads the authors 
to believe that such forms are definitely rare if not abnormal. This obser- 



126 A MANUAL OF THE ASPERGILLI 

vation is, in effect, confirmed by George Smith (1931). From textiles in 
particular he has isolated many small-spored strains but none with large 
spores. Possibly the present collection contains as many large-spored 
strains as it does because the authors have regarded them as curiosities, 
and for that reason retained them, whereas scores of strains of such com- 
mon species as A . repens or A . amstelodami have been isolated and forthwith 
discarded. 

In contrast to the small-spored forms, where complete duplication be- 
tween large numbers of isolates is the rule, among the large-spored forms 
there is a marked tendency for each strain to present a somewhat different 
cultural picture, which is commonly coupled with differences in morphology. 
This would suggest that these forms are unstable and variable, but such a 
conclusion is refuted by their behavior in culture. To illustrate, culture 
NRRL No. 131, a strain of Aspergillus echinulatus, has for 20 years of con- 
tinuous culture by the authors retained its distinguishing characters, simi- 
larly the single known strain of A. medius has been under observation in 
this and European laboratories for more than 40 years without appreciable 
change. 

Thus the problem of assigning a relatively small number of quite distinct 
strains is presented. To describe each of them would merely add to the 
confusion already existing, hence they have been grouped somewhat, choos- 
ing either historic cultures that have become widely distributed or cultures 
of marked individuality as representing specific names. Homogeneity 
among the strains brought together is not claimed. The names A. glaucus 
and E. herbariorum are not identified with particular organisms in this dis- 
cussion. 

In setting apart a so-called "large-spored series" the authors do not, in 
any sense, wish to imply close relationship or genetic continuity within this 
subgroup. Species are grouped together primarily as a matter of con- 
venience, and (E.) herbariorum is selected as the series designation primarily 
because of Man gin's usage of this species name to cover all of the large- 
spored forms. 

Key 

A. Conidial heads green. 

1. Asci ripening within 2 to 4 weeks. 

a. Ascospores 6.5 to 7.5ju in long axis A . mangini n. comb. 

b. Ascospores 7.5 to 8.5ju in long axis A. umbrosus Bain, and Sart. 

c. Ascospores 9.0 to lO.Oju in long axis 

A. echinulatus (Delacr.) Thorn and Church 

2. Asci ripening slowly, 2 to 3 months, colonies favored by 40 per cent sugar. 

a. Ascospores with equatorial ridges and furrow A. medius Meiss. 

b. Ascospores usually without equatorial ridges and furrow 

A. carnoyi (Biourge) Thorn and Raper 

B. Conidial heads white A. niveo-glaucus Thorn and Raper 



THE ASPERGILLUS GLAUCUS GROUP 127 

Aspergillus mangini (Mangin) n. comb. 

Synonyms : Eurotium herbariorum ser. minor Mangin, Ann. des Sci. 
Nat., Bot. (ser. 9) 10: 365. 1909. 
Aspergillus minor (Mangin) Thorn and Raper, U. S. D. A. 
Misc. Publ. No. 426, p. 27. 1941. 

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted, 
attaining a diameter of only 1 to 2 cm. in 3 weeks, irregular and wrinkled, 
cream colored to bluish-brown, conidial heads present or lacking, small 
perithecia present or lacking, mostly abortive; reverse uncolored to orange- 
maroon. 

Colonies upon Czapek's solution agar with 20 percent of sucrose plane 
or somewhat wrinkled in the central area, spreading evenly, attaining a 
diameter of 8 to 10 cm. in 3 weeks (fig. 33 A), predominantly brick-red in 
color becoming maroon in age, perithecia abundant and borne in a close 
felt of red-encrusted hyphae at the agar surface, conidial heads few in 
number, projecting above the perithecial layer, generally distributed over 
the entire colony, but occasionally concentrated in localized areas; reverse 
in shades of deep red-brown. 

Perithecia abundant, largely embedded in and obscured by a close 
mycelial felt at the agar surface, yellow to orange, globose to subglobose, 
mostly 100 to 120m in diameter, occasionally up to 150m; asci 14 to 16m; 
ascospores lenticular, commonly 6.6 to 7.4m by 5.2 to 5.8m occasionally up 
to 7.8m in long axis, finely roughened in the equatorial area, ridges low and 
rounded or pyramidal in section, furrow generally definite, shallow but 
often steep-sided, V-shaped. 

Conidial heads few, generally scattered, projecting above the perithecial 
layer, pale blue-green in color, radiate, mostly 150 to 200m in diameter, but 
frequently larger; conidiophores smooth, pale to dark brown, mostly 700 
to 800m in length, occasionally reaching 1 mm. broadening to 15 to 18m 
below the vesicular apex; vesicles subglobose, 30 to 40m; sterigmata in a 
single series, 8 to 10m by 4 to 5m; conidia dull green, elliptical to sub- 
globose mostly 6.0 to 7.5m and frequently 8.0m in long axis. 

Represented by culture NRRL No. 117 isolated from an unpainted 
board, as type ; and by several additional cultures isolated in this laboratory. 
Culture NRRL No. 115, received in 1937 from Oscar W. Richards, of the 
Spencer Lens Company, differs from the type by producing ascospores of 
somewhat smaller size and with less evident furrow and ridges. Thus, it 
may represent a strain transitional between Aspergillus mangini and A. 
ruber. However, it is not sufficiently different from A. mangini, either 
culturally or morphologically, to warrant its description as a separate 
species or as a distinct variety. 

Mangin (1909), in his study of the group, found specimens in his collec- 




Fig. 33. Comparative growth of large-spored members of the Aspergillus glaucus 
group upon 20 percent sucrose Czapek agar at room temperature : A , A . mangini, NRRL 
No. 117, 4 weeks. B, A. umbrosus, NRRL No. 120, 4 weeks. C, A. echinulatus, 
NRRL No. 131, 6 weeks. D, A. niveoglaucus, NRRL No. 127, 4 weeks. E, A. medius, 
NRRL No. 125, 6 weeks. F, A. carnoyi, NRRL No. 126, 4 months. (Reprinted 
from Thorn and Raper, "The Aspergillus glaucus Group," U.S.D.A. Misc. Pub. 426: 
1-46. 1941.) 

12S 



THE ASPERGILLUS GLAUCUS GROUP 129 

tion with ascospores less than 7.5/x in long axis, yet larger than those of the 
small -spored forms which he described. Having no faith in any of the 
descriptions existing at the time, he called the aggregate Eurotium her- 
bariorum series minor. The cultures cited above have sufficient common 
characters to warrant the belief that they are variants of a common stock 
which may be constituted a species aggregate to which we apply the 
name Aspergillus mangini. 

Aspergillus umbrosus Bain, and Sart., in Soc. Mycol. de France, Bui. 

Trimest. 28: 267-269, pi. XII. 1912. 

Probable synonyms : A . mutabilis Bain, and Sart., in Soc. Mycol. de 

France, Bui. Trimest. 27: 458, pi. XVII. 1911. 
A. mollis Bain, and Sart., Soc. Mycol. de France, 
Bui. Trimest. 27: 453, pi. XVI. 1911. 

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted, 
attaining a diameter of 0.5 to 1.0 cm. in 3 weeks, raised, tufted, white to 
orange-red, bearing neither perithecia nor conidial heads ; reverse colorless 
to orange-brown. 

Colonies upon Czapek's solution agar with 20 percent of sucrose, plane 
or somewhat wrinkled, spreading evenly or irregularly (fig. 33 B), reaching 
a diameter of 8 to 10 cm. in 3 weeks, predominantly vinaceous red to orange- 
brown in color, consisting largely of a surface felt of sterile hyphae encrusted 
with orange-red granules enmeshing abundant perithecia, occasionally 
characterized by a loose floccose overgrowth bearing scattered perithecia, 
conidial heads pale blue-green, widely scattered and projecting above the 
perithecial layer; reverse in red-brown shades. 

Perithecia abundant, yellow to orange, globose to subglobose, largely 
embedded in a felt of sterile red-encrusted hyphae at the agar sur- 
face, occasionally borne in a loose aerial felt, mostly 120 to 140m in 
diameter, rarely up to 175m; asci 14 to 16m; ascospores lenticular, mostly 
7.2 to 8.0m by 5.6 to 6.4m, occasional spores up to 8.4m in long axis, finely 
roughened to smooth in the equatorial areas, ridges low and generally 
rounded, furrow shallow, commonly V-shaped (fig. 34 A); conidial heads 
few, scattered, projecting above the perithecial layer, pale bluish-green, 
radiate, compact, mostly 175 to 250m in diameter; conidiophores smooth, 
colorless to brownish, 700 to 850m in length, broadening to 15 to 20m 
below the expanded, domelike vesicular apex; vesicle 25 to 40m in diameter; 
sterigmata in a single series, 10 to 12m by 4.5 to 6m; conidia pale green, 
elliptical to subglobose, spinulose, mostly 7 to 8m in long axis, frequently 
larger. 

Represented in the present collection by culture XRRL No. 120 received 
in 1925 from Dr. Florence A. McCormick, by European strains, and by 
several cultures isolated by the authors. 



130 



A MANUAL OF THE ASPERGILLI 




Fig. 34. Ascospores representative of A, A. umbrosus, and B, A. echinulatus. In 
each species upper left, right, and center spores represent surface profile views; 
lower left, optical section in profile; lower center, surface in face view; and lower 
right, optical section in face view. (Reprinted from Thorn and Raper, "The Asper- 
gillus glaucus Group," U. S. D. A. Misc. Pub. 426: 1-46. 1941.) 



THE ASPERGILLUS GLAUCUS GROUP 131 

Culture XRRL Xo. 123 contributed by Dr. Paul Simonart as an un- 
named culture from the Biourge collection differs from the species as above 
described by consistently producing ascospores with walls entirely smooth, 
whereas in other characters the ascospores duplicate essentially those of 
XRRL Xo. 120. Further XRRL Xo. 123 produces colonies of lighter color 
than other strains under observation and may, in fact, represent a fungus 
comparable to that described as Aspergillus mutabilis by Bainier and 
Sartory. 

Aspergillus u?nbrosus, A. mutabilis, and A. mollis were described by 
Bainier and Sartory (1911c, 1912b) primarily upon the basis of colony color 
(pigment production) and conidial apparatus, with the ascopsores of A. 
umbrosus recorded as slightly less in long axis (8.0 by 5.6m) than those 
of the other species (8.4 by 5.6m)- After careful consideration of the 
three descriptions and detailed study of the strains in the authors' posses- 
sion showing in general the ascospore described by Bainier and Sartory, 
it is believed that they had at hand three cultural variants of the same 
species. A . umbrosus is retained as the species designation, as it is believed 
that their description of this species more adequately pictures the cultural 
and morphological characters of the fungi under consideration than either 
of the earlier descriptions, which are left as probable synonyms. 

Aspergillus echinulatus (Delacr.) Thorn and Church, The Aspergilli, 

p. 107. 1926. 

Synonyms : Eurotium echinulatum Delacr., Soc. Mycol. de France, Bui. 

Trimest. 9: 266, pi. XIV, fig. III. 1893. 
.4. brunneus Delacr., in Bui. Soc. Mycol. France 9: 185, PL 

XI, fig. 9, 1893; was described as the conidial stage. 
E. verruculosum Vuill., Soc. Mycol. de France, Bui. Trimest. 

34:83. 1918. 

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted, 
1.5 to 3.0 cm. in diameter after 4 weeks, marginal area blue-green from 
conidial heads and central portion reddish-brown from an overgrowth of 
sterile encrusted hyphae, perithecia lacking; reverse deep orange. 

Colonies upon Czapek's solution agar with 20 percent of sucrose slow- 
growing, plane or somewhat wrinkled, spreading irregularly, attaining a 
diameter of 7 to 8 cm. in 4 weeks (fig. 33 C), commonly mottled in appear- 
ance due to the uneven distribution of green conidial heads above the 
underlying orange-red perithecial layer; conidial heads bottle-green, 
abundant, commonly crowded in localized areas but scattered thinly 
throughout the remainder of the colony; perithecia abundant and borne 
in a felt of hyphae encrusted with red granules at the agar surface, con- 
spicuous where not obscured by massed green heads ; reverse cinnamon to 
deep red-brown. 



132 A MANUAL OF THE ASPERGILLI 

Perithecia abundant, embedded in a looose felt of sterile red hyphae at 
the agar surface (fig. 26 E), yellow, globose to subglobose, mostly 125 to 
150/x in diameter, and occasionally up to 175m; asci 18 to 22m; ascospores 
lenticular, mostly 9 to 10m by 6.5 to 7.5m, occasionally up to 11m in long 
axis, conspicuously roughened in the equatorial area, furrow pronounced, 
broad, ridges prominent and irregular (fig. 34 B). 

Conidial heads densely crowded hi localized areas and scattered through- 
out the remainder of the colony, bottle-green in color, radiate, consisting of 
relatively few, long, divergent chains of conidia, commonly 250 to 300m 
in diameter but often larger or smaller; conidiophores smooth-walled, 
colorless to brown shades, commonly 700 to 850m in length, occasionally in 
excess of 1 mm., broadening from 5 to 7/z at the base to 15 to 20 /x below the 
vesicular apex; vesicle 25 to 35/x in diameter, consisting of a domelike 
terminus of the broadening conidiophore ; sterigmata in a single series, not 
crowded, bottle-shaped, 12 to 15/x by 5 to 7m; conidia elliptical, pyriform, 
or subglobose, echinulate, mostly 8 to 10m hi long axis, commonly larger or 
smaller, extremely variable. 

Represented by NRRL No. 131 isolated in 1921 from figs received from 
California. A subculture of this strain, forwarded by Miss Margaret 
Church about 1926, is maintained in the Centraalbureau ; the two lines re- 
main identical. No ascosporic stage has in the authors' experience been 
found in culture NRRL No. 133 received in 1937 from George Smith as 
A. echinulatus Delac, and obtained by him from Biourge, but its conidial 
development duplicates NRRL No. 131 and it is apparently correctly as- 
signed. Da Fonseca's and the Centraalbureau 's isolations of A. echinu- 
latus maintained at Baarn produce somewhat smaller ascospores (8 to 9m 
by 6.2 to 7.0m) and conidia than No. 131, but otherwise agree essentially 
with the species description as given above. Somewhat further reduction 
in ascospore size is seen in cultures NRRL No. 523 isolated from honey, 
and NRRL No. 137 received from George Smith and Raistrick as Asper- 
gillus mongolicus Biourge (nomen nudum). Although these are less red 
in color than No. 131 and appear distinct in culture, the authors do not 
feel warranted in separating them as a species or variety, believing that they 
represent only variations from the general type designated as A. echinulatus. 

Bainier and Sartory described A. disjunclus (1911b) and A. repandus 
(1911c) as vigorous species possessing ascospores 11 by 6m and 11.2 by 
5 6m, respectively. Authentic cultures of these species are not now avail- 
able, but the descriptions as published would seem to place them close to 
A. echinulatus. 

Probable Synonyms 

Several Aspergilli with ascospores ranging near that described for A. 
echimdatus appear in the literature. Unfortunately, the details of asco- 



THE ASPERGILLUS GLAUCUS GROUP 133 

spore markings are not given and there is a dearth of data to identify them. 
Some of these names are given : 

A. disjunctus Bainier and Sartory, in Soc. Mycol. France Bui. 27: 346-368. PI. X, 
XI. 1911. Ascospores described as 11.2 by 5.6m with furrow and crests. 

.4. repandus Bainier and Sartory, in Soc. Mycol. France Bui. 27: 463. PI. XVIII. 
1911. Ascospores described as 11 by 6m with furrow but no crests. 

.4. mencieri Sartory and Flament, in Compt. Rend. Soc. Biol. (Paris) 83: 1114-1115. 
1920. Ascospores 10 by 4.7m with furrow and crests. 

A. godfrini Sartory and Roederer, in Assn. Frangaise pour l'Avancement des 
Sciences, 42nd Session, Tunis 1913. pp. 601-603. 1914. Conidial stage only. This 
was growing at blood heat and warmer. Its general description suggests affinity 
with the large-spored species of the A. glaucus group. It has not been seen in cul- 
ture by us. 

Strains with the particular measurements reported for the thiee asco- 
sporic species listed above have not come into our collection, yet presum- 
ably they and many more with minor variations may be found. 

Aspergillus medius Meiss., in Bot., Ztg. 55: (337)-344, (353)-357. 1897. 

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted, 
0.5 to 1.5 cm. diameter in 6 weeks, tufted, consisting of a dense growth of 
yellow-brown hyphae bearing neither conidial heads nor perithecia ; reverse 
in shades of yellow-brown. 

Colonies upon Czapek's solution agar with 20 percent of sucrose at room 
temperature very slow-growing (optimum 20°C. ±), strongly wrinkled, 
tufted, irregular in outline, attaining a diameter of 5 to 6 cm. in 6 weeks 
(fig. 33 E), colony center deep orange-red becoming yellow to white at the 
margin, which is characterized by bundles (becoming branching columns 
at lower temperatures) of hyphae bearing dark-green conidial heads in the 
manner of loose divergent coremia; perithecia ripening very slowly, ma- 
turing ascospores in 2 to 3 months, mostly abortive; conidial heads rela- 
tively few (more abundant and larger at lower temperatures) and borne 
either in coremiform masses or scattered throughout the colony; reverse in 
shades of orange -maroon. 

Perithecia scattered, mostly abortive, borne in a dense felt of orange-red 
hyphae, very slowly ripening, globose to very irregular in form, extremely 
variable in size, rarely attaining a diameter of 125m, containing very few 
mature ascospores; asci 18 to 20m; ascopsores sparingly produced, lenticu- 
lar, mostly 8.8 to 9.6m by 6.0 to 6.8m, occasionally 10m in long axis, some- 
what roughened in the equatorial region, furrow broad and shallow, ridges 
prominent, relatively thin and irregular. 

Conidial heads deep-green, radiate, compact, and of two types: Small 
heads 100 to 150m in diameter borne on loose coremiform columns, and 
larger heads 200 to 250m in diameter often scattered throughout the colony, 



134 A MANUAL OF THE ASPERGILLI 

produced more abundantly at 12° to 15° C. than at room temperature; 
conidiophores colorless to brown, mostly 250 to 35(V in length, enlarging 
to 15 to 20ju below the vesicle; vesicle subglobose, mostly 30 to 40/i in 
diameter; sterigmata in a single series, crowded, short, 7 to 8/t by 4 to 5/x; 
conidia green, globose to subglobose, finely echinulate, thick-walled, mostly 
8 to 10/x in diameter, but frequently larger or smaller. 

Represented in the NRRL collection by culture No. 124 which was re- 
ceived in 1924 from Raistrick, who in turn received it from the Centraal- 
bureau. It is believed to be Meissner's original strain (1897). Subcultures 
of this strain are currently maintained at Baarn and by George Smith in 
London. The three lines remain identical as shown by parallel cultures 
during recent study. 

This fungus is distinguished particularly by (1) its very slow growth 
upon 20-percent sucrose Czapek agar at room temperature, (2) its tardiness 
in producing perithecia and especially in ripening ascospores, (3) its sparse 
production of ascospores, and (4) its formation of aerial hyphal bundles 
bearing conidial heads in loose coremiform fashion. Further, it grows 
much more rapidly upon Czapek agar containing 40 percent of sucrose than 
upon that containing 20 percent, a difference in concentration which does 
not materially affect the growth rate of such vigorous species as A. repens 
and A. chevalieri. Growth is much more rapid at 20° C. than at 28° to 
30° C. (fig. 12). The fungus attains a more favorable form at the lower 
temperature, at which there is a heavier growth of mycelium, a more 
extensive development of aerial hyphal columns, and a greater production 
of conidial heads and perithecia. 

Culturally this fungus is easily separated. from all other species of the 
group, except possibly A. carnoyi. 

Aspergillus carnoyi (Biourge) Thorn and Raper, IT. S. Dept. Agr. Misc. 

Pub. 42G, p. 34. 1941. 

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted 
reaching a diameter of only 3 to 4 mm. in 6 to 8 weeks, thin, white, bearing 
neither conidial heads nor perithecia; reverse colorless. 

Colonies upon Czapek's solution agar with 20 percent of sucrose at room 
temperature extremely slow growing (optimum 18° to 20° C), reaching 
a diameter of 7 to 8 cm. in 6 to 8 weeks, irregular in outline, somewhat 
floccose, forming a deep felt, bearing abundant perithecia and scattered 
conidial heads (fig. 33 F), orange-brown in central area to orange at margin 
from abundant perithecia in a loose network of sterile hyphae encrusted 
with orange-red granules; reverse in orange-red shades. 

Perithecia late in developing, abundant, yellow to orange, globose to 
subglobose, mostly 125 to 175/* in diameter but frequently larger or smaller, 
borne in a loose floccose felt of sterile brown hyphae; asci 16 to 18ju, 



THE ASPERGILLUS GLAUCUS GROUP 135 

typically 8-spored, frequently with some or all spores aborted; ascospores 
lenticular, variable in size and pattern, mostly 7.2 to 8.4/z by 6 to 6.5^ but 
often larger (up to 9.0/x in long axis) or smaller (down to 6.5yu in long axis), 
generally smooth-walled but occasionally roughened in equatorial area, 
generally rounded but often flattened and occasionally indented, with 
ridges wholly absent or indefinite, and with furrow absent or present as a 
trace only. 

Conidial heads sparsely produced, commonly scattered, dull gray-green, 
radiate, compact, mostly 150 to 200/z but often up to 250/x in diameter; 
conidiophores smooth-walled, colorless, long, commonly up to 2 mm. in 
length, uniform in diameter, 12 to 18m, to just below the vesicle; vesicle 
subglobose, 40 to 50^ in diameter and occasionally larger; sterigmata in a 
single series, crowded, bottle-shaped, 10 to 12^ by 5 to 6m; conidia globose 
to subglobose, echinulate, dull green, mostly 8 to 10^. 

Species description based upon culture NRRL No. 126 received in 1937 
as Aspergillus carnoyi Biourge from George Smith and by him earlier from 
Biourge. Presumably the culture is type, although Biourge 's description 
of the species remains unpublished. The culture is distinct, not only differ- 
ing in its colony character and in the length of its conidiophores but espe- 
cially in the variable character of its ascospores. The majority of spores, 
although much larger, resemble those of A. repens, whereas spores with 
rough walls are occasionally produced. This culture is, therefore, somewhat 
of an exception to the general rule of constancy in ascospore pattern, and 
the variability of its spores affords one of the best characters for its identifi- 
cation. 

Aspergillus niveo-glaucus Thorn and Raper, U. S. Dept. Agr. Misc. Pub. 

426, p. 35. 1941. 
Synonyms: vl. glaucus mut. alba Bloch., Deut. Bot. Gesell. Ber. 50: 
248-256. 1932. 
A. glaucus var. albida. Speg., An. del Mus. Nac. de Buenos 
Aires 6: 332. 1899. 

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted, 
1 cm. in diameter after 4 weeks, white to cream, bearing abundant small 
conidial heads but no perithecia; reverse colorless to yellow-brown. 

Colonies upon Czapek's solution agar with 20 percent of sucrose slow 
growing, plane, spreading irregularly, 6 to 8 cm. in diameter after 4 weeks 
(fig. 33 D), thinning toward the margin, with mycelium in shades of yellow- 
orange becoming cinnamon brown in age, more or less obscured by abun- 
dant white heads, and with perithecia abundant, yellow (PI. Ill F), em- 
bedded and irregularly clustered in a close felt at the agar surface ; reverse 
yellow at margin to deep brown at colony center. 

Perithecia abundant, yellow, globose, to subglobose, mostly 100 to 125/x 



136 



A MANUAL OF THE ASPERGILLI 



in diameter, occasionally larger, commonly clustered, borne in an inter- 
rupted surface felt of buff to brown hyphae; asci 15 to 17/z in diameter; 
ascospores lenticular, mostly 7.2 to 7.8/i by 5.0 to 5.6/i, smooth-walled 



« 




i 



3 it I 






• 



I 







t%> a 



\ 






B 










^:D 



Fig. 35. Aspergillus glaucus group, conidial heads. A and B, A. niveo-glaucus, 
NRRL No. 127: A, surface view showing loose, radiate character of heads, X 35; 
B, photomicrograph of the same showing large, globose vesicle fertile over almost 
the entire surface, X 600. C and D, Aspergillus restrictus, NRRL No. 154: C, surface 
view showing long, columnar heads, X 120; D, photomicrograph of the same showing 
characteristic small vesicle fertile on the uppermost surface only, X 600. 

except in equatorial area, furrows broad and shallow, ridges prominent, 
roughened, rounded or acute and often appearing ragged. Conidial heads 
abundant, white (fig. 35 A), often becoming browned in age, radiate, mostly 
250 to 300 /x in diameter; conidiophores smooth-walled, colorless to brown, 



THE ASPERGILLUS GLAUCUS GROUP 137 

mostly 1,000 to 1,500m, rarely longer, broadening to 16 to 20// below the 
vesicle; vesicle subglobose, 40 to 50/x in diameter (fig. 35 B); sterigmata in 
a single series, crowded, 8 to 10/x by 3 to 4m; conidia elliptical to pyriform, 
colorless, spinulose, 6 to 8m in long axis. 

The cultural description is based upon NRRL No. 127 of this collection 
as type; this is duplicated by XRRL No. 130 received from Baarn in 1938 
as Aspergillus glaucus Link mut. alba Bloch. Culture NRRL No. 128, 
received in 1935 from George Smith and Raistrick and by them from 
Biourge as .4. albidus Speg., differs from type by consistently producing a 
greater quantity of conidial heads which are commonly of smaller size; 
the ascosporic stage of the two is indistinguishable. Apparently the name 
"albidus" is a manuscript use in which Spegazzini's variety (1899) has been 
raised to species rank, presumably by Biourge. Blochwitz' mutation albus 
assigned to .4. glaucus is untenable, because no definite organism can be 
designated as A. glaucus. Furthermore, the specific name albus applied to 
an Aspergillus has already been used in another section of the genus. 

It is possible that Blochwitz (1932c) is right in regarding this as a muta- 
tion, but there is nothing to indicate which particular large-spored form is 
the parent species. The strain maintains its identity in culture and hence 
must be regarded as a species. Yuill (1939), in contrast, has described 
white mutants of A. nidulans and A . fumigatus and has properly designated 
them as mutants, for they appeared in cultures under observation and are 
known to have been derived from typical chromogenic strains. 

THE ASPERGILLUS RESTRICTUS SERIES 

Thorn and Church (1926) called this series the "Intermediate Forms" 
between the A. glaucus and the A. fumigatus groups. George Smith, wish- 
ing to emphasize the resemblance of the conidial apparatus to the monover- 
ticillate Penicillia called the lot the A. penicilloides group (1931), thus 
suggesting Spegazzini's species as the typical member. However, the 
important relationship indicated by the structure of the conidial apparatus 
is not with Penicillium but with the A. glaucus group of which these forms 
appear to be merely reduced members. All of these forms, like many of 
the A. glaucus forms, grow characteristically under conditions of physiolog- 
ical drought — represented by their frequency upon mildewed textiles as 
studied by Smith (1928) and Galloway (1930) or in concentrated cane 
products as reported by Owen (1923). Similarly we have found them in 
many situations in which physiological drought is attained by physical 
dryness or osmotic concentration attained by the presence of high per- 
centage of sugar or sodium chloride. 

This natural relationship is on the whole better indicated by accepting 
Smith's .4. restrictus as typical, and regarding the other known members 



138 A MANUAL OF THE ASPERGILLI 

of this series as allied species. To perpetuate an assignment to a subdivi- 
sion entitled the Microaspergilli, as suggested by some authors, would com- 
plicate nomenclature without compensating values. The series is, there- 
fore, keyed as a part of the great A. glaucus group. 

Series diagnosis: Perithecia not found. Colonies growing weakly 
or restrictedly upon Czapek's solution agar, more freely upon wort agar, 
especially well on high concentrations of sugar or salt; green, dark green, 
grayish-green, to brownish-green in various strains and under varying 
conditions; surface growth consisting of conidiophores only, or of mycelial 
felts more or less buckled or heaped; conidiophores smooth, slender, more 
or less sinuous, septate; vesicles vary from convex or lens-like areas on the 
broadened apices of conidiophores to definitely ovate to globose enlarge- 
ments, fertile over all or the upper fraction of such surfaces; heads mostly 
definitely columnar, less commonly radiate, hemispherical or almost glo- 
bose, especially when young; sterigmata in one series, mostly closely packed 
over the fertile area, varying from 2 to 3m by 5 to 6m up to 3 to 4/x by 6 to 
10m; conidia barrel-shaped to ovate, mostly in dark greenish shades, smooth 
or slowly becoming echinulate or roughened as in the A. glaucus group, 
commonly adherent into long chains which are packed into columns. 

Series Key 

A. Conidiophores broadening upward to produce a convex vesicular apex varying 

from 8 to 20m in diameter, producing a long slender column of conidia 

A. gracilis Bainier 

B. Conidiophores broadening more abruptly, forming a more definite vesicular area. 

1. Vesicles more convex, toward hemispherical. 

a. Slime development evident A. conicus Bloch. 

b. Slime absent or not conspicuous A. restrictus G. Smith 

2. Vesicles ovate to hemispherical, columns of conidial chains more or less 

conspicuous A . penicilloides Speg. 

Differences between the above may appear to be of somewhat minor 
character. Nevertheless, each of the sections accounts for sufficient litera- 
ture to necessitate separate consideration . 

Aspergillus gracilis Bainier, in Bui. Soc. Myc. France, 23: 92, pi. IX, figs. 

11-14. 1907. 

Colonies on Czapek's solution agar very slow growing, reaching a diam- 
eter of only a few millimeters in several weeks (fig. 36A), variously plane 
or convoluted or buckled with close textured mycelium at first white, then 
slowly green to very dark green, with radiating lines of vegetative mycelium 
about the denser area of the colony, growing somewhat better upon wort 
agar and upon Czapek's agar containing high concentrations of sugar, re- 
verse in yellowish shades, conidial heads in columns up to 200 or 300m long 



THE ASPERGILLUS GLAUCUS GROUP 



139 



by 10 to 20 even 25m in diameter, straight or twisted. Conidiophores 
mostly arise as very short branches of aerial hyphae up to 20 to 30/z long or 
less commonly up to 100 to 125m even up to 250m and gradually broadening 
toward the apex to a vesicular area which is very flat, dome-like, almost the 
effect of a truncated cone, 8, 10 to 20m or more in diameter, bearing a single 
series of sterigmata, 6, 8, 10 by 2 to 3m, or in particular strains growing out 
into little conidiophores producing secondary heads ; conidia at first barrel 
form then subglobose about 3m in long axis in Bainier's strain, progressively 
larger in related strains. No perithecia found. 

The type culture has not been seen. Forms with approximately the 
morphology described, however, have appeared in strains NRRL No. 145 
(Thorn 4246) from moldy corn; Thorn 4197.3 (culture lost) from Owen in 




Fig. 36. A, Aspergillus gracilis, NRRL No. 145, on Czapek's solution agar with 
20 percent sucrose, after incubation for 2 weeks at room temperature. B, A. 
restrictus, NRRL No. 154, on Czapek's solution agar with 20 percent sucrose after 
18 days at room temperature. 



the sugar laboratory, New Orleans; and a strain from Thaxter appearing as 
a contaminant in a Papulaspora culture. A. gracilis may thus be assumed 
to represent a form occasionally found especially in very concentrated 
substrata. Biourge later contributed a culture, as type, of A. hypo- 
janthinus originally described by him as Penicillium hypojanthinum Biourge 
(1923). Three other cultures from Biourge labeled P. (Microaspergillus) 
hickeyi, Microaspergillus albo-marginatus, and P. (M.) guegueni (figured in 
his monograph, Plate XX, but not described) appear in culture to be only 
minor variations of this general form. Later, Biourge sent his undescribed 
A . sartoryi. This grew more freely and showed the conidiophore and vesicle 
of A. gracilis; conidia were 6 to 7m or greater in long axis, definitely rough 
and corresponded almost exactly with a culture received from George Smith 



140 A MANUAL OF THE ASPERGILL1 

as A. gracilis (XRRL No. 156). Whether further accumulation of strains 
will justify giving Biourge's proposed name, A. sartoryi, sectional or varietal 
status or merely emphasize the completeness of the series as showing great 
variations in structural detail is left uncertain. There does not appear to 
be any warrant for preserving A. gracilis var. exiguus Bainier and Sartory 
(1912 a). According to the description this variety differs slightly in phy- 
siological characters from A . gracilis Bainier. 

A. conicus Blochwitz, in Dale, Ann. Mycol. 12: 38. 1914. Previously 
described by Dale as a Penicillium in Ann. Mycol. 10: 465. 1912. 

See also Thorn and Church "The Aspergilli" p. 125. 1926. 

Th? outstanding character of this species is found by microscopic ex- 
amination of colonies which become buckled or contorted from a close 
felting of mycelia. Relationship back toward the A . glaucus group is found 
in the sterigmata, and in the elliptical conidia. Heads differing little in 
microscopic structure from A. gracilis are found to be more or less 
completely submerged in dark green to almost black slime. Many strains 
have been collected from widely separated regions showing all gradations 
from a trace of slime only to complete submergence of the heads, 
particularly in old cultures. 

Dale isolated the strain first described from English soil, and sent dupli- 
cate cultures labeled Penicillium sp. to Blochwitz and to Thorn; Thom re- 
turned the very brief descriptive note without name as published by Dale 
in 1912. Later Blochwitz proposed by letter to her the name only, A. 
conicus, which was published by Dale in 1914 without further description. 
Later, in his own publication, Die Gattung Aspergillus (1929), Blochwitz 
denies the slime development as a character of his organism and redescribes 
the species in terms to make A . conicus cover the section of this group re- 
presented by Smith's A. restrictus (1931). Since the name was already in 
the literature for the slimy series which certainly appears in culture from 
widely separated places, it should stand as originally applied. Whether 
the slime disappears in some strains long kept on artificial media is not 
settled. 

In 1928, Biourge contributed under the manuscript name A. cyanogenes 
a strain which reproduced the characters given by Thom and Church for A . 
conicus. This or a nearly related organism appeared as a contaminant in 
several of the cultures received from Biourge. The culture (Thom No. 
4733.138) figured in his Monograph (1923) as Xo. 126 in Plate XXI, under 
P. glabrum Dale was another. Still another strain from Biourge, labeled 
A. viridans differed only in the delayed development of the slimy covering 
of the columnar mass of conidia. 




Plate IV 

A (upper left), Aspergillus restrietus Smith, NRRL No. 154. B (upper right), Aspergillus fumigatus 
Presenilis, NRRL No. 163. C (center left), Aspergillus nidulans (Eidami Wint., XRRL No. 192. D (center 
right), Aspergillus rariecolor (Berk, and Br.) Thorn and Raper, NRRL No. 195-1. E (lower left), Aspergillus 
ustus (Bain.) Thorn and Church, NRRL No. 278. F (lower right), Aspergillus flaripes (.Bain, and Sart.) 
Thorn and Church, NRRL No. 295. Figure .4 growing upon Czapek's solution agar containing 20 percent 
sucrose and 0.5 percent peptone; all other cultures growing upon standard Czapek's solution agar. (Color 
photographs by Haines, Northern Regional Research Laboratory. Reproduced through co-operation of 
Chas. Pfizer & Co., Inc.) 



THE ASPERGILLUS GLAUCUS GROUP 141 

Neill in his study of the Aspergilli of New Zealand (1939) applied the 
name A. caesiellus Saito (1904) to what was manifestly some strain near A. 
conicus Blochwitz, and then placed the whole group in that species. 

A. restrictus G. Smith, in Jour. Text. Inst. 22: IT 15, fig. V. 1931. 
Species characterization by George Smith 

Colonies growing very poorly on Czapek agar; growing moderately well 
on wort agar, dark dull green, gradually turning grey or brownish -grey; 
reverse in some cultures uncoloured, in others green to dark green ; surface 
velvety at first, becoming wrinkled and often acquiring a warted appearance 
(PI. IV A and fig. 36 B); heads forming long, compact, slender columns 
(fig. 35 C) up to 350m by 20 to 30m in diameter; conidiophores arising mostly 
from substratum but also as branches of aerial hyphae, commonly 50 
to 100m, occasionally 150-200^ long by 3 to 3.5m in diameter, often with one 
or two septa, smooth, sinuous, uncoloured; vesicles flask-shaped 7.5 to 14/i 
in diameter; sterigmata in one series, borne on upper surface of vesicles 
only, 6 to 9m by 2.5 to 3m (fig. 28 C); conidia rough, spinulose, elliptical or 
somewhat pyriform, often showing a distinct connective, dark greenish- 
brown, 4 to 6.5m by 3 to 4m, mostly 4.5 to 6m by 3 to 3.5m; perithecia not 
found. The young conidia are hyaline and cylindrical and almost appear 
to be segments of enormously elongated septate sterigmata. They 
gradually swell without increasing in length, at the same time becoming 
pigmented, but even in old heads they adhere strongly together in columns 
of parallel chains ; and mounts (fig. 35 D) made in lactophenol usually show 
compact, twisted, columnar masses of ripe conidia, both attached to and 
separated from the heads. 

It is evident from Smith's discussion of a large accumulation of cultures, 
especially from the textile industry, that the usual colony-type in his ex- 
perience is not the "conicus" type with its slime but the columnar head 
described as A. restrictus and accepted here as giving the most appropriate 
name to the series. 

Among organisms belonging to this series, Smith isolated a single strain 
which differed from typical A. restrictus in "showing somewhat larger di- 
mensions throughout and, more particularly, in the production of conidia 
up to 10 m or more in length," and to which he assigned the designation 
Aspergillus restrictus var. B. (G. Smith, Jour. Text. Inst. 22: T115. Figs. 
IV, VI, and VIII. 1931). Upon examination by us, this culture (NRRL 
No. 148) was found to correspond fairly closely to his published description. 
However, we do not believe it is sufficiently distinct to warrant continued 
separation as a variety since other strains showing intermediate dimensions 
arc encountered. 



142 A MANUAL OF THE ASPERGILLI 

Aspergillus penicilloides Spegazzini, in Rev. Agrar. Veter. 
La Plata, p. 246. 1896. 

Spegazzini 's description was emended by Thorn and Church (The As- 
pergilli, p. 126, 1926) then broadened by Smith (Jour. Text. Inst. 22, pp. 
114-115, 1931) as follows: 

"Colonies growing fairly slowly on wort agar, rich dark green with paler 
edge, turning darker and duller, and finally becoming dirty greenish-grey 
overgrown with sterile hyphae; reverse brown, greenish-brown, and dark 
green in patches; surface much wrinkled and folded; heads globose when 
young, 40 to 70 /x in diameter, becoming columnar, somewhat ragged, and 
up to 200^ long; conidiophores arising either from substratum or from aerial 
hyphae, smooth, thin-walled, 75 to 150 m long by 6 to 10 n in diameter; 
vesicles rather sharply marked off from conidiophores, pear-shaped to sub- 
globose, 15 to 23/xin diameter, fertile over the upper half or two-thirds; 
sterigmata in one series, crowded, 8 to 10 n by 2.5 to 3.5 n; conidia ovate, 
barrel shaped or nearly spherical, usually showing connective, rough, 3.5 
to 5m by 3.2 to 4/x, with very dark colored walls." 

Thorn and Church had strain No. 4197.3 isolated from cane products in 
Louisiana by Owen and agreed to by Spegazzini; NRRL No. 151 (Thorn 
No. 7), also from cane products, fits this description satisfactorily; as did 
also Biourge's strain labeled A. pertardus. Smith reports various strains 
from mildewed textiles. 

It would thus appear that the vesicular area in the series varies from the 
curved apex of a clavate conidiophore as in A. gracilis, to a fairly well- 
defined hemispherical vesicle as in A. restrictus, and finally to an almost 
globose body as in A . penicilloides. All have so much in common with each 
other and with the A. glaucus group that their relationship as "degraded" 
mutants appears probable. 

Aspergillus itaconicus Kinoshita, in Botan. Mag. Tokyo 45: 60-61. 1931. 

Probable synonym: A. varians Wehmer, in Bot. Centralb. 80: 460-1. 

1899; also in Wehmer Monogr. 77-79, Taf. I, fig. 
1. 1899-1901. 

Diagnosis from Kinoshita's organism obtained from Dr. Westerdijk. 
Colonies on Czapek's solution agar forming dense felts 1 to 2 mm. deep, 
white or yellowish, ridged and irregular, with scattered long-stalked green 
heads upon dry areas and on the glass ; fruiting more abundantly upon malt 
agar, and particularly upon Czapek's solution agar containing 20 percent 
sugar; reverse of colony and agar yellow to orange-reddish upon some 
media; heads large, light green, globose to radiate, breaking up easily under 
the coverglass; conidiophores smooth, colorless, 8 to 16 ju or larger in diam- 
eter and up to several millimeters in length under some conditions, with 



THE ASPERGILLUS GLAUCUS GROUP 143 

walls 0.5 to 1.5m in thickness and splitting lengthwise as A. niger 
when broken; vesicles 15 to 4G>, globose or subglobose (fig. 28, D); sterie- 
matain 1 series 8 to 9 /x by 1.5 to 2 p\ conidiamore or less pyriform, 4.3 to 5m 
by 3.5 to 4/x, finely echinulate, of the A . glaucus type. 

Diagnosis is drawn from culture NRRL No. 161 (No. 5344 of Thorn) re- 
ceived from Westerdijk as the type of Kinoshita's A. itaconicus and agrees 
closely with the description given by Wehmer for A . varians (cf . Thorn and 
Church, The Aspergilli, p. 127, 1926), not with Thorn and Church's descrip- 
tion of their culture No. 115 which was subsequently lost. A. itaconicus 
is so closely related in its physiological responses and in several of its struc- 
tural characters to the A. glaucus group that it is placed as an ex- 
treme variant at the end of this whole group. 

Kinoshita described his organism as a producer of itaconic acid (1931a) 
and detailed his experimental work (1931b). The culture as distributed to 
laboratories outside of Japan seems to conform to Kinoshita's description 
and to produce the acid, but in quantities too small for commercial develop- 
ment. More recently, Calam, Oxford, and Raistrick (1939) have recovered 
itaconic acid as a metabolic product of A. terreus. From an industrial 
point of view, this source appears to be far more promising. 

VARIATION IN THE ASPERGILLUS GLAUCUS GROUP 

The genetic history of the Aspergilli is an untouched field. Separation 
into large groups is easily made definite enough to include all but a few 
strains. Within these groups, variation is so great that differentiation of 
species requires critical examination and comparison of material, including 
extensive culture. Unwilling to undertake this, Neill (1939) disposed of 
the whole group with yellow perithecia by calling them all A. glaucus. On 
the other hand, Mangin (1909), with the same problem of variability before 
him, found the ascospores sufficiently distinctive and dependable to warrant 
proposing to separate A . amstelodami and A . chevalieri as separate species 
in the A. glaucus group, leaving certain aggregates admittedly inadequately 
studied. Bainier and Sartory (1911b, 1911c, 1912), working with members 
of these ill-defined species, used color production as the basis for separation. 
They cited ascospore measurements as incidental details in description. 
Because they appear to have had only a few strains in culture and to have 
described them all as new species, the task presented few difficulties to 
them; but unfortunately without the original cultures, no one has ever been 
able to identify their species with confidence. Raistrick and his colleagues 
(1934, 1937, 1938, 1939), using cultures named as received and including 
an unpublished series from Biourge, have given quantitiative figures as to 
pigment production and pigment mixtures for each culture listed by the 
name on the tube, without reporting comparative study of the morphology 
found. 



144 A MANUAL OF THE ASPERGILLI 

It is clear that wide mycelial or colony variations may be found in nature 
between strains that retain the ascospore characters of the species. Several 
such groups have been held by the authors for 30 years or more and fur- 
nish convincing evidence that Mangin was justified in using the ascospore 
as the stable and readily determinable integrating character. Some of these 
forms retain colony characters in fairly stable form through many transfers 
on many laboratory substrata and over a period of years. Others grown in 
various substrata in Petri-dish cultures show sectors or other irregularities 
in colony habit, which can be picked out and established as strain variants 
that maintain their special characteristics in continuous culture. 

The possibility that variants of similar nature might be induced by chemi- 
cal stimulation led Thorn and Steinberg (1939) to select from the authors' 
collection certain strains that had remained fairly constant for many years. 
Of this group they subjected strain NRRL No. 90 of A. amstelodami, found 
apparently stable for 30 years, to extensive chemical stimulation (1940a). 
These experiments yielded two groups of effects: (1) A progressive reduc- 
tion in the production of conidial heads and of perithecia, and (2) a great 
increase in the mass of vegetative mycelia. In no case was spore produc- 
tion completely suppressed although reduced to inconspicuous quantity. 
The conidia and ascopsores when examined were found to have retained 
the size and markings characteristic for the species, whereas the mass of 
vegetative mycelium became excessive and formed a floccose or cottony 
mass entirely different in colony appearance from the original. 

At this point they reversed the procedure and applied stimulants designed 
to reestablish spore production (1940b). As a result, the final cultures 
show abundant green heads with normal conidia and numerous perithecia 
with ascospores retaining the characters of the species. In routine labora- 
tory examination these extreme variants would not suggest the original 
strain of A . amstelodami, although both types of variants produce conidia 
and ascospores typical of the species. 

In 1928, Barnes studied the possibility of heat in inducing variations. 
He used a strain reported as "Eurotium herbariorum (Wigg) Link," which 
had been isolated and maintained in his laboratory for several years without 
apparent changes. Unfortunately, no description of his normal strain was 
given, but a strain received from Westerdijk as "Barnes' normal strain" 
proves to be identical with A. amstelodami (Baarn strain, NRRL No. 89; 
or strain No. 90 as used by Thorn and Steinberg). No reasons were offered 
for the original identification. 

Barnes described a series of experiments in which the spores of his or- 
ganism were subjected to heat under varied conditions, then planted. 
From the resulting colonies he described 11 variants, 6 of which are avail- 
able in the Centraalbureau. The authors' transfers of these have been 



THE ASPERGILLUS GLAUCUS GROUP 145 

checked against the descriptions in Barnes' paper and obviously represent 
his isolations. 

In the following list, his designations appear as quotations, followed by 
our identifications based upon careful cultural study: 

"Flame" variant is A. ruber. 

"Green flame" is A. repens. 

"Blue conidial" is .4. chevalieri var. intermedins. 

"Creamy" is a pale yellowish strain of Yuills' genus Cladosarum. 

"D Brown" is A. uslus. 

"C Yellow" is A. amstelodami. 
Of these, "C Yellow" (NRRL No. 112) shows the ascosporic pattern and 
differs little from Barnes' normal strain or the authors' A. amstelodami. 
"Flame" (NRRL No. 59), "Green flame" (culture discarded), and "Blue 
conidial" (NRRL No. 85) show ascospores of the glaucus group but differ 
markedly in pattern. Among large numbers of induced variations, changes 
in the characters of the ascospore have not been found during this study. 
"D Brown" (culture discarded) produces no ascospores but develops the 
hiille cells and conidial heads characteristic of the Aspergillus ustus group. 
These four forms belong to ubiquitous species quite abundant as con- 
taminants where plant material is handled. Such contamination is not 
satisfactorily excluded by the work reported. 

"Creamy" (NRRL No. 143) presents a different problem. The possi- 
bility that this " Cladosarum" was actually derived from the "normal" A. 
amstelodami is not excluded. Proliferation of the sterigmata in the head of 
Aspergilli, especially among the A. glaucus lot, is very common. The 
branches produced sometimes are found sterile but usually become diminu- 
tive conidiophores with very small vesicles and groups of sterigmata pro- 
ducing normal spores. The Yuills' Cladosarum olivaceum (NRRL No. 374) 
was found as a conspicuous variant or contaminant in their culture of A. 
niger (1938). The colony, conidiophore, vesicle, primary sterigmata, and 
initial secondary sterigmata are produced as in A. niger, then instead of 
chains of conidia with the newest or youngest conidia at the bases of the 
chains and connected directly with the sterigmata, chains of cells are pro- 
duced that replicate the sterigmata; occasionally a chain is interrupted by 
one cell producing a group of new chains, thus acting as a primary sterigma. 
These chains of cells lengthen not at the base as in Aspergillus but at the 
distal end. In spite of prolonged search, which shows that the cells toward 
the outer ends of such chains lose definiteness as sterigmata, the authors 
cannot confirm the finding of a single terminal conidium on each chain as 
reported by the Yuills. This morphological picture is repeated by Barnes' 
"Creamy" strain, which must therefore be interpreted in terms of Yuills' 
genus. Only the two isolations are known thus far. Their failure to pro- 



14G A MANUAL OF THE ASPERGILLI 

duce true spores and their rapid loss of vitality observed in the present cul- 
tures lead to the hypothesis that they are both variants from Aspergillus 
and belong to the "monster" type of organisms that fail to survive in com- 
petitive environments. Rare occurrences of the "monster" type such as 
these can hardly be regarded as permanent members of the fungous flora, 
even though the individual can be kept viable by regular vegetative trans- 
fer. It is doubted, therefore, whether generic designation is warranted. 

Occurrence and Economic Importance 

The members of the A . glaucus group are among the most common molds 
on earth. Th°y are extremely abundant in nature, and live under all sorts 
of conditions, thriving in moist and dry situations. However, they are most 
conspicuous upon concentrated substrata, such as drying plant products of 
all kinds as they come from the field and reach storage just above the ab- 
solute low percentage of water for stability. They are equally com- 
monplace in sweetened products, including jams, jellies, soft sugars, honey, 
soft candies; in salted products such as meats, pickles, etc.; in dried foods 
where preservation is a complex of sugars and acids; upon manufactured 
leather goods exposed to humid conditions ; upon clothing and textiles stored 
in moist atmospheres; and upon soft wood inadequately cured or subjected 
to improper storage. Nevertheless they are frequently overlooked in rou- 
tine culture because they grow poorly upon substrata commonly employed 
and soon become overgrown by bacteria and more rapidly developing molds. 
In following the deterioration of products by cultural procedures, these 
forms will appear with amazing frequency if care is taken to select media 
approximating the osmotic concentration of the substance examined. 

Molds of this group herald incipient spoilage. As excess water increases, 
first other molds, then bacteria, appear and complicate the decomposition 
process. The presence of any of these organisms is evidence of the earliest 
stages of decomposition, and is generally followed by the invasion of other 
and more destructive molds and bacteria. Insofar as is known, products 
infected by these forms in pure culture are non-poisonous; but naturally 
occurring materials in which they appear, or even predominate, should be 
considered "suspect" because of the possible presence of toxigenic forms. 

Pathogenicity 

Members of the A . glaucus group have been reported in connection with 
various types of ailments of man and domestic animals. A. hageni of 
Hallier (1870) and A. repens have been occasionally reported as fruiting in 
the external canal of the human ear; A. montevidensis Talice and MacKin- 
non (1931) was isolated from a case of otomycosis ; A . mencieri Sartory and 
Flament (1920) was described from sputum of a consumptive; Fonseca 



THE ASPERGILLUS GLAUCUS GROUP 147 

(1930) isolated, in Brazil, A. amstelodami from a case of mycetoma of the 
foot; and .4. keratitis Ball was described as on the cornea (Thorn and 
Church, The Aspergilli, p. 86, 1926). There is thus data enough to justify 
belief that an occasional strain of the A . glaucus group is found in connec- 
tion with lesions in man. We know nothing of the manner of infection and 
have no evidence that these organisms appear in any consistent manner as 
pathogens. Quevedo's case in which A. maydis (1912) appeared as a cause 
of poisoning of horses reads plausibly; this, however, is rendered doubtful 
by the observations of other investigators who have reported in widely 
separated places such masses of mycelium and spores without encountering 
similar injury. A . fontoynonti of Gueguen (1909 and 191 1) isolated from an 
abcess was not found pathogenic in subsequent animal experiments. 
Species which grow well at blood heat have possibilities of pathogenesis; 
fortunately, however, most members of the group do not grow at 37° C. 



Chapter X 
THE ASPERGILLUS FUMIGATUS GROUP 

Outstanding Characters 

Conidial heads columnar, in shades of green through dark green to 

fuliginous. 
Vesicles flask-shaped, typically fertile over the upper half. 
Sterigmata in one series, crowded. 

Conidiophores smooth-walled, usually colored in shades of green. 
Conidia globose, echinulate, green. 

Working with lung material from birds dying oi aspergillosis. Presenilis, 
about 1850, described and figured the species Aspergillus fumigatus to well 
that there has never been any doubt as to the morphology of the mold 
present. The investigator of molds in culture, however, quickly finds that 
this type of conidiophore and head characterizes not a single pathogenic 
strain but a multitude of variant forms that are abundant in soil, upon 
decaying vegetation, and. in fact, wherever organic materials are under- 
going even the slightest aerobic decomposition. To all oi these forms, col- 
lectively, the designation Aspergillus fumigatus group is applied. . 

The group falls naturally into two series: 

Cultures strictly conidial, varying from velvety to floccose I. fumigatus series 

Cultures producing perithecia and ascospores, conidial development generally 
limited A. fischeri series 

THE ASPERGILLUS FUMIGATUS SERIES 

Aspergillus fumigatus Presenilis, in Beitrage sur Mykologie. p. 81, pi. 10, 

tigs. 1-11. Frankfurt, 1850 53. Thorn and Church. 

The Aspergilli. p. 120. 1926. 

Colonies upon Czapek's solution agar spreading broadly over the sub- 
stratum, in some strains strictly velvety (PI. IV B and fig. 37 A.), in others 
more or less floccose with varying amounts of tufted-aerial mycelium to deep 
felted or extremely floccose forms (fig. 37 B), white at first, becoming green 
with the development of heads but varying considerably in the final shade of 
green, often becoming dark green to almost black in age. Reverse and 
substratum, in some strains uneolored. in others showing varying amounts 
of yellow, or again passing over in dark red shades in age. Conidial heads 
columnar, compact, varying in measurement from strain to strain up to 400 
by 50m, hut usually much shorter, occasionally very small. Conidiophores 

14> 










■f ' 



Fig. 37. Aspergillus fumigatus group. A-C, A. fumigatus: A, Typical, heavy 
sponng strain, XRRL No. 178, on Czapek's solution agar, 10 days, room temperature; 
B, Floccose, light sporing strain of same species, XRRL Xo. 171 ; and C, Photomicro- 
graph of typical conidial heads showing characteristic form of vesicles and crowded 
stengmata in a single series, X 500. D-F, Aspergillus fischeri: D, Strain XRRL 
Xo. 186, growing upon Czapek's solution agar, characterized by very abundant 
perithecia and few conidial heads; E, Portion of a colony enlarged showing crowded 
penthecia more or less obscured by loose enveloping hyphae, X 6.0; and F, Asci con- 
taining ascospores at various stages of maturity, X 500. 

149 



150 A MANUAL OF THE ASPERGILLI 

short, smooth, usually densely crowded, up to 300m (in occasional strains 
up to 500^) m length by 2 to 8m in diameter, frequently more or less green 
colored, especially in the upper part, arising directly from submerged 
hyphae or as very short branches from aerial hyphae, septate or unseptate, 
gradually enlarging upward and passing almost imperceptibly into the 
apical flask-shaped vesicles. Vesicles up to 20 to 30m in diameter, usually 
fertile on the upper half only (fig. 37 C). Sterigmata in one series, usually 
about 6 to 8m (varying from 5 to 10m) by 2 to 3m, crowded, closely packed 
with axes roughly parallel to the axis of the conidiophore. Conidia dark 
green in mass, echinulate, globose, mostly 2.5 to 3m in diameter with ex- 
tremes ranging from 2 to 3.5m. Sclerotia or perithecia are not found. The 
species grows well at temperatures up to 45° C. or even higher, and is com- 
monly present in compost and other material undergoing decomposition at 
high temperatures. 

The species description presented is a composite rather than an exact 
citation of detailed data about one strain, but NRRL No. 163 (Thom No. 
118) may be considered typical. Organisms coming within this series as 
described are world-wide in distribution and omnivorous in habit. They 
are regularly abundant in soil and in decomposing organic masses ; they are 
recoverable from apparently sound cereals, corn, oats, wheat, etc. ; they are 
encountered as pathogenes in the air passages of birds and occasionally as 
lung parasites of mammals, including man. One strain (NRRL No. 164) 
was named A. cellulosae by Hopffe (1919) because of its ability to break 
down cellulose, but when examined, it presented no morphologic differences 
from hundreds of common isolates from soil. 

Efforts to induce perithecium formation by the conidial strains of A.fumi- 
gatus have been disappointing. Organisms maintained in culture over long 
periods and subjected to multitudes of transfers upon all sorts of substrata 
have failed to give any response suggesting perithecium formation. 

Extremes of variation in different strains range from colonies character- 
ized by crowded conidiophores rising vertically from submerged hyphae to 
a height of 300m, or perhaps at times 500m, then producing columns of co- 
nidia sometimes up to 400 by 50m, to very floccose forms in which spore for- 
mation is generally retarded and in which conidiophores develop as very 
short branches of aerial hyphae and produce short columnar heads. Many 
of these variants can be isolated and maintained in culture, thus they have 
been made the types of species by earlier workers. Others encountered 
upon unique substrata have been given specific names in the belief that the 
substratum relation was obligate. Yuill (1939) isolated a buff-colored 
mutant from a typical green strain and applied to it the designation A. 
fumigatus var. helvola (fig. 17 C). Shortly thereafter Steinberg and Thom 
(1940) likewise recovered an essentially uncolored mutant from a typical 
green strain. Both forms still remain unchanged in culture. 



THE ASPERGILLUS FUMIGATUS GROUP 151 

If the taxonomist could seize a half dozen widely spaced variants and 
destroy those which bridge the gaps between, identification as separate 
species would be easy. This, however, becomes impossible when large 
numbers of isolates are cultivated and studied in comparative culture. In 
attempting to divide this great series into tangible entities, earlier workers 
have created a long list of species and, while we do not consider these to be 
valid, they are presented in alphabetical order with the places of description. 
It is believed that all of these should be regarded as synonyms of A . fumi- 
gatus Fres. 

A. aviarius Peck, in N. Y. State Museum Rept. 44, p. 25, pi. 4, figs. 9-12. 1891- 

A. bronchialis Blumentritt, in Ber. Deutsch. Bot. Ges. 19: 442-446, PI 22 ; figs 1-A- 
1901; also ibid. 23: 419-427, PI. 19, figs. 1, 3, 6, 7, 8, 19, 23. 1909. 

A. calyptratus Oudemans, in Arch. Neerl. Ser. II. 7: 283. Tab. XIII. 1902. 

A. cellulosae Hopffe, in Centralb. f. Bakt. etc., Abt. 83: 531-537. 1919. 

A. desseyi Spegazzini, in Physis (Rev. Soc. Argentina Cien. Nat.) VIII: 115-117, 
1 figure. 1925; review only seen, Rev. Appd. Mycol. 4: 542. 1925. 

A.fumigatus var. alpha Sion and Alexandrescu, in Compt. Rend. Soc. Biol. (Paris) 
64: 288-289. 1908. 

A.fumigatus var. minimus Sartory, in Bui. Acad. Med. Paris 3 Ser. 82: 304-305. 
1919. 

A. fumigatus var. lumescens Blumentritt, in Ber. Deut. Bot. Ges. 23: 419-427, 
PL 19, figs. 5, 6, 18-21. 1905. 

A. glaucoides Spring, in Bull. Acad. Sci. Belg. 19: 560-572. 1852. 

A. lignieresi Cost, et Lucet, in Ann. Sci. Nat. Ser. 9, II: 119, tab. 5, fig. 18-23. 
1905. 

A. nigrescens Robin, in Histoire Naturelle des Vegetaux Parasites, p. 518, Paris. 
1853. 

A. pulmonum hominis Welcker, in Kuchenmeisters Parasiten II, p. 144. This is 
discussed and figured by Theodor von Dusch, in Virchow's Archiv. (n. f. 1) 11: 561- 
566. 1857, but no ground is given for separating it from A.fumigatus. 

A. ramosus Hallier, in Zeitschr. Parsit. 2: 266-269, pi. 6, figs. 1-6. 1870. 

A. syncephalis Gueguen, in Les Champignons parasites de l'homme et des animaux 
299 pp., 12 pi. Paris, 1904. 

A. virido-griseus Cost, and Lucet, in Ann. Sci. Nat. Bot. IX. 2: 140. 1905. 

THE ASPERGILLUS FISCHERI SERIES 

Aspergillus fischeri Wehmer, in Centralb. f. Bakt., etc., 2 abt., 18: 

390-2, figs. 1-5. 1907. 

Synonyms:^!, fumigatus — ascosporic, see Thorn and Church, in Am. 
Jour. Bot. 5: 91-92. 1918. See also Thorn and Church, 
The Aspergilli, p. 132. 1926. 
Sartorya fumigata Vuillemin, in Compt. Rend. Acad. Sci. 
(Paris) 184, No. 3: 136-137. 1927. 

The conidial form is strikingly similar to Aspergillus fumigatus. Colonies 
grow well upon Czapek's solution agar with conidial heads sparingly pro- 



152 A MANUAL OF THE ASPERGILLI 

duced at room temperature (fig. 37 D), but more abundantly at 37° C. 
Conidial heads frequently small and generally of a lighter green color than 
those of typical A. fumigatus. Perithecia quickly and abundantly pro- 
duced in 'most strains dominating the colony appearance (fig. 37 E), 
commonly up to 300 M in diameter, not colored, or very pale salmon, with 
walls scarcely colored, consisting of a single layer of cells, crushing easily, 
covered by a loose network of uncolored sterile hyphae. Asci abundant, 




Fig. 38. Ascospores of Aspergillus fischeri NRRL No 181 (- Thorn No 4651 2) 
Upper left, center, and right represent surface, P^.^f^je^fiSKon 
section in profile; lower center, surface in face view; and louer right, optical section 



in face view. 



8-spored, filling the perithecium within a few days, 8 to 10/x by 10 to 12/z 
subglobose (fig. 37 F), breaking down quickly to leave the perithecium full 
of ripe ascospores. Ascospores biconvex, uncolored, usually about 7 by 4/x, 
consisting of a central body 5 by 4 M , with two frilled equatorial banks about 
1 M in width, roughened with echinulations or anastomosing bands on each 
convex surface (fig. 38), separating into two valves in germination. 

Culture NRRL No. 181 (Thorn No. 4651.2), received as type from 
Wehmer in 1923, is apparently identical with numerous isolations rom 
American sources. Many strains have been seen, including a series from 
sputum of human cases showing lung involvement by X-ray examination. 



THE ASPERGILLUS FUMIGATUS GROUP 153 

It is regarded as world-wide in distribution but seemingly not abundant 
anywhere. The species grows well at temperatures of 37° C. and higher. 
Some additional perithecial forms have been described with characters 
suggesting relationship with A. fischeri. However, these are inadequately 
known in culture and assignment here must remain somewhat provisional. 

A. rnalignus Lindt, in Arch. Exp. Path. Pharmakol. 25: 257-271, fig. 1-11. 1889. 

Lindt probably had before him some strain which corresponded closely with 
Wehmer's Aspergillus fischeri despite the fact that his description of the conidial 
apparatus offers sufficient contrast as to lead to question. 

A.fumigatoides Bainier and Sartory, in Bull. Soc. Myc. France 25: 112, pi. 5. 1909. 

While the describers believed this strain close to A. fumigatus, Thorn and Raper 
(1941) found it necessary to place the organism studied under this name by Gould 
and Raistrick (1934) in A. Pseudoglaucus, although the original description and 
figures of Bainier and Sartory probably represented material close to, or identical 
with, A. fischeri. 

Sartorya fumigaia (Fres.) Vuill., in Compt. Rend. Acad. Sci. (Paris), 184(3): 136- 
137. 1927. 

Sartory, Sartory, and Meyer (1926) reported that a culture of A. fumigatus sub- 
jected to radiation produced an ascosporic form. Later this was designated by 
Vuillemin (1927) as a new genus Sartorya. Neither Vuillemin, nor Sartory, Sartory, 
and Meyer appear to have known the relation between A. fumigatus and A. fischeri 
although it was pointed out by Thorn and Church in 1918. Since the material named 
Sartorya does not appear to have been distributed or fully described, Sartorya fumi- 
gate (Fres.) Vuillemin may be regarded as a synonym for A. fischeri Wehmer and the 
generic name dropped. 

It is believed probable that Aspergillus fischeri Wehmer represents the 
primary organism of this group and that from it the conidial form A. fumi- 
gatus developed as a species lacking entirely the ascosporic phase. The 
fact that A . fumigatus was described first merely reflects the much greater 
abundance of this species. 

Occurrence and Economic Importance 

Aspergillus fumigatus is an extremely cosmopolitan mold and occurs with 
particular frequency in soil containing appreciable organic materials, upon 
vegetable matter undergoing slow decomposition, and upon imperfectly 
dried, stored grains. The mold is an important agent in many decomposi- 
tion processes, particularly at temperatures above 37° C. Growing success- 
fully at 45° to 50° C, within the lower reaches of thermophilic decomposi- 
tion, it is able to operate within a range where most fungi are excluded. 
Whereas some forms have been described as very active agents of decom- 
position (e.g. A. cellulosae of Hopffe, 1919), their more significant role is 
believed that of forerunners of active bacterial decomposition on the one 
hand, and as slow destroyers of more resistant tissues on the other. Asper- 
gillus fischeri, though much less abundant, may be found in situations 
generally similar to those yielding A. fumigatus. 



154 A MANUAL OF THE ASPERGILLI 

Pathogenesis 

There is an extensive pathological literature which covers the occurrence 
of A. fumigatus in lesions of birds and mammals, including man, together 
with biochemical and animal experimentation. Such experiments have 
repeatedly proved that the organism is pathogenic to fowls confined in con- 
gested quarters in which moldy grain, straw, and other plant remains are 
abundant. Direct inoculation to the cornea in laboratory animals causes 
lesions characteristic for the species. 

Infection of human beings occasionally appears, and observations seem 
to indicate that the patients generally have been exposed to air carrying 
large numbers of spores. Allergists have reported asthmatic conditions 
arising from sensitization to this species, and Bern ton (1930) reports having 
successfully treated a patient by means of an extract prepared from the 
spores and mycelium of A. fumigatus. The occurrence of A. fischeri in 
cases grouped with A. fumigatus is clear indication that the pathogenic 
principle, whatever it is, is generally present in the group although it may 
vary in its intensity among different strains as indicated by workers such 
as Costantin and Lucet (1905). 

Among organisms known to be, or believed to have been, pathogenic 
strains of A. fumigatus, the following named forms may be cited: A. 
gratioti, A. malignus, A. fumigatoides, A. virido-griseus, A. bronchialis, A. 
glaucoides, A. nigrescens, A. pulmonum hominis, A. ramosus, and A. avia- 
rius (see p. 151). For a more complete discussion of this group in relation 
to disease in birds and mammals, the reader is referred to Thorn and 
Church's The Aspergilli (1926) and Dodge's Medical Mycology (1935). 

Antibiosis 

Anslow and Raistrick (1938a) reported the production by Aspergillus 
fumigatus of a substance to which they applied the name, fumigatin, and in 
the same year (1938b) reported the species to produce a second metabolic 
product termed spinulosin, which they had previously isolated from Peni- 
cillium spinulosum. In 1942 fumigatin was further discussed by Oxford 
and Raistrick as a powerful agent against such bacteria as Bacillus anthra- 
cis, Escherichia coli, Salmonella typhi-murinum, Staphylococcus albus, S. 
aureus, Streptococcus viridans, and Vibrio cholorae. Also in 1942 Waksman, 
Horning, and Spencer reported an antibiotic substance, termed fumigacin, 
to be produced by A. fumigatus, and presented methods of differentiating 
this from fumigatin as studied by Raistrick and associates. Fumigacin was 
found to be both bactericidal and bacteriostatic in its action and to be 
effective in fairly high dilutions in inhibiting the growth of gram -positive 
cocci and bacilli ; it was much less effective against the gram-negative mem- 
bers of the coli-aerogenes group. Both fumigatin and fumigacin have been 
found toxic to experimental animals. 



Chapter XI 
THE ASPERGILLUS NIDULANS GROUP* 

Outstanding Characters 

Conidial heads short columnar, usually dark green with primary and 
secondary sterigmata. 

Conidiophores smooth- walled, more or less browned, usually sinuate, 
commonly less than 200m long and terminating in dome-like or hemi- 
spherical vesicles. 

Conidia globose, echinulate, 3 to 4m in diame'ter. 

Perithecia usually present; ascospores purple-red in color and character- 
ized by equatorial bands. 

Large, thick-walled, globose bodies, termed "hiille cells" (by Eidam), 
forming an irregular layer around the perithecia. 

Aspergillus (Sterigmatocyslis) nidulans was described by Eidam in 1883. 
Since that time the general type of organism covered by his diagnosis has 
become fairly well-known and certain striking characters have become 
recognized as defining a number of cosmopolitan strains or species, com- 
monly referred to as constituting the Aspergillus nidulans group. 

The hiille cells of Eidam, first described in A. nidulans, appear also in 

various transformations in the A. versicolor, A. ustus, and A . flavipes groups, 

together with other common characters indicative of close relationship. 

They do not appear in the A . clavatus, A . glaucus, or A . fumigatus groups 

which are characterized by single sterigmata, nor do they occur in the 

sclerotium forming groups, A. candidus, A. niger, A. wentii, A. tamarii, A. 

flavus, and A. ochraceus. 

Group Key 
I. Ascospores present. 

A. Ascospores smooth-walled. 

1 . Equatorial ridges two in number. 

a. Ridges 0.5 to 1.0m wide, margin entire. 

Conidial heads green A. nidulans (Eidam) Wint. 

Conidial heads white A. nidulans mut. alba Yuill 

b. Ridges 1.5 to 1.8m wide, margin entire 

A. nidulans var. latus Thom and Raper 

c. Ridges 3.0 to 4.9m wide, margin dissected, starlike 

A. variecolor (Berk, and Br.) Thom and Raper 

2. Equatorial ridges usually four in number 

A. quadrilineatus Thom and Raper 

B. Ascospores rough-walled A. rugulosus Thom and Raper 

1 Abridged from: Thom and Raper, The Aspergillus nidulans group, Mycologia, 
Vol. XXXI, No. 6, 653-669, Nov.-Dec. 1939. 

155 



156 A MANUAL OF THE ASPERGILLI 

II. Ascospores lacking. 

A. Hiille cells forming irregular masses, suggestive of sclerotia 

A. caespitosus Raper and Thorn 

B. Hiille cells absent; heavy walled sterile hyphae present 

A. unguis (Emile-Weil and Gaudin) Thorn and Raper 

Aspergillus nidulans (Eidam) Wint. in Rab. Krypt.-Fl. I 2 : 62. 1884. 

Synonyms : Sterigmatocystis nidulans Eidam in Cohn, Beitr. Biol. Pflan- 
zen 3 : 392-41 1. pi. 20-22. 1883. 
Diplostephanus nidulans (Eidam) Langeron, Compt. Rend. 
Soc. Biol. Paris, 87: 343-345. 1922. 

Colonies upon Czapek's solution agar plane, spreading broadly, dark 
cress green (Ridgway, PI. XXXI) from abundant conidial heads during the 
first two weeks (PI. IA, IVC, and fig. 41 A) ; perithecia developing fron the 
center of the colony outward after the first few days, separately produced, 
often abundant (PI. IVC) ; sectoring occasional; reverse of colony in varying 
shades of purplish-red during the growing period, becoming very dark in 
age. Heads short, columnar, ranging from 40 to 80m by 25 to 40m, com- 
monly 60 to 70m by 30 to 35m (PI- IB and fig. 4 C); conidiophores com- 
monly sinuous, with walls smooth, in shades of cinnamon brown (PI. IC), 
ranging from 60 to 130m, commonly 75 to 100m in length, about 2.5 to 3m 
near the foot, increasing to 3.5 to 5m below the hemispherical vesicle (fig. 
39 A); vesicle 8 to 10m in diameter; sterigmata in two series, primary 5 to 
6m by 2 to 3m and secondary 5 to 6m by 2 to 2.5m; conidia globose, rugulose, 
3 to 3.5m in diameter, green in mass. 

Perithecia developed separately within or upon the conidial layer (PI. 
IA), globose, ranging from 100 to 175m in diameter, commonly 125 to 150m, 
with outer layer a yellowish to cinnamon colored envelope of scattered 
hyphae bearing hiille cells up to 25m in diameter; wall composed of one 
layer of cells, dark reddish-purple; in ripening becoming a mass of 8-spored 
asci which break down quickly leaving the ascospores free. Ascospores 
purple-red, lenticular, smooth-walled with 2 equatorial crests (PI. ID and 
fig. 43 A), spore bodies about 3.8 to 4.5m in length by 3.5 to 4m in breadth, 
equatorial crests pleated with margin sinuous and entire ranging from 0.5 
to 1m in width (Table 1). 

Diagnosis based primarily upon culture XRRL Xo. 187 (Thom Xo. 
4640.5) obtained from the Bainier collection in Paris. Other strains as- 
signed to Eidam 's species included many isolations from American soil and 
decaying vegetation, as well as cultures from European contributors. 
Common. 

The range of ascospore measurements found in a representative group 
of cultures is shown in the accompanying table. 

In assigning Eidam's species name to the members of this series it is 
obvious that there are discrepancies. He described the perithecium as 



THE ASPERGILLUS NIDULANS GROUP 



157 







Fig. 39. Conidial structure in the Aspergillus nidulans group, X 900: A u Typical 
conidiai head of A. nidulans, XRRL Xo. 187; Ao, Diminutive head of the same strain; 
B, Typical head of .4. caespitosus, XRRL Xo. 1929. 




Fig. 40. Aspergillus nidulans. A, Conidial heads, X 370. Conidiophores arise 
either from the substratum or from aerial hyphae. B, Hiille cells, X 740. 








Fig. 41. Aspergillus nidulans group; different species growing upon Czapek's 
solution agar at room temperature. A, A. nidulans NRRL No. 194, typical strain 
producing very abundant dark green conidial heads. B, Naturally occurring muta- 
tion characterized by white heads, isolated from the preceding strain by Edward 
Yuill and described as A. nidulans mut. alba. C, A. rugulosus NRRL No. 207, 
characterized by heavy perithecium production, an almost complete absence of 
conidial structures, and a tendency of colonies to split in central areas as shown. 
D, A. variecolor NRRL No. 1954, characterized by the production of abundant large 
perithecia. E, A. unguis NRRL No. 216, characterized by an absence of perithecia 
and hulle cells. F, A. caespitosus NRRL No. 1929, characterized by the production 
of masses of hiille cells suggesting abortive perithecia. 



158 



THE ASPERGILLUS NIDULANS GROUP 



159 



having a firm almost sclerotioid wall, whereas the wall is found to contain 
but one laj^er of cells. He figured the asci as few and scattered in a mycelial 
matrix within which ascospore production occupied many weeks. One 
isolated strain in our collection produces asci in this manner. For it, the 
varietal name, A. nidulans var. latus, was proposed by Thorn and Raper 
(1939) on account of very broad crests on the ascospore in contrast to the 
usual types in A. nidulans which come much more closely to those indicated 
in Eidam's figures. 

TABLE 1 
Ascospore variation in strains of Aspergillus nidulans 



Culture number 



NRRL 193.. 
NRRL 188. . 
NRRL 189.. 
NRRL 194.. 
NRRL 195*. 
NRRL 191 . . 
NRRL 187.. 
NRRL 192.. 
NRRL 198. . 
NRRL 199.. 



Overall dimension of spores 



6.2-6.6 
6.0-6.6 
6.0-6.4 
5.4-5.8 
5.4-5.8 
5.4-5.8 
5.4-5.8 
5.0-5.4 
4.8-5.4 
4.8-5.2 



x 3. 6-3. 8 M 
x 3. 6-3. 9/x 
x 3. 6-3. 8m 
x 3. 6-3. 8m 
x 3. 6-3. 8m 
x 3. 6-3. 9m 
x 3. 6-3. 8m 
x 3. 6-3. 8m 
x 3. 6-3. 8m 
x 3. 6-3. 8m 



Width of crests 


1.0m ± 


1.0m ± 


1.0m ± 


0.6-0. 8m 


0.6-0. 8 m 


0.6-0. 8m 


0.7-0. 8m 


0.5-0. 6m 


0.5-0. 6m 


0.5m ± 



Dimensions of spore bodies 

4.2-4.6 x 3. 6-3. 8m 
4.0-4.6 x 3. 6-3. 9m 
4.0-4.4 x 3. 6-3. 8m 
4.0-4.4 x 3. 6-3. 8m 
4.0-4.4 x 3. 6-3. 8m 
4.0-4.4 x 3. 6-3. 9m 
3.9-4.3 x 3. 6-3. 8m 
4.0-4.4 x 3. 6-3. 8m 
3.8-4.4 x 3. 6-3. 8m 
3.8-4.2 x 3.6-3. 8m 



* White mutant from culture NRRL No. 194; described by Yuill as Aspergillus 
nidulans mut. alba. 

A. nidulans mut. alba Yuill, in Jour, of Botany (London) 

175, pi. 618. 1939. 

Colonies of the variety on Czapek's solution agar differ from the species 
in the entire absence of green color (fig. 41 B). The ascospores have the 
characters of the species. 

Culture obtained by Yuill as a mutant from a normal green strain of A. 
nidulans under investigation in his laboratory. Our record number is 
NRRL 195; the normal and parent strain is NRRL 194. 

Aspergillus nidulans var. latus Thorn and Raper. Mycologia 31 : 657. 1939. 

Colonies on Czapek's solution agar differing from the species in colony 
development characterized by a felt of predominantly sterile mycelium; 
few conidial heads; fairly abundant perithecia developed in the mycelial felt 
and each surrounded by a thick covering of hlille cells (fig. 42 C), very slowly 
ripening and containing few and scattered asci in abundant sterile mycelium. 
Ascospore bodies smooth-walled, purple-red, 3.8 to 4.5m by 3.5 to 4m, with 
crests 1.5 to 1.8m in width. 

Type culture NRRL No. 200 received from the Centraalbureau in 1909 
and remaining constant in culture since that time. Its antecedent history 
is not known. 



160 A MANUAL OF THE ASPP^RGILLI 

Aspergillus quadrilincatus Thorn and Raper. Mycologia 31: 6G0, figs. 2- 

4. 1939. 

Colonies on Czapek's solution agar spreading, plane or slightly wrinkled, 
with tendency toward floccosity, central area gray with a definite purplish 
tinge, and olive-green conidial areas toward the margin, occasionally as 
sectors; perithecia developing separately but abundantly throughout the 
colony; reverse purplish-red; heads short columnar, green, mostly 60 to 70/z 
by 30 to 35m, occasionally larger or smaller; conidiophores sinuate, smooth- 
walled, dull brownish in color, 50 to 75/i in length by 3.5 to 4.5m wide, 
broadenng to 7.5 to 9/x at the hemispherical vesicular areas; primary 
sterigmata 5 to 6/x by 2 to 3m, secondary sterigmata 5 to 7 m by 2 to 2.5m; 
conidia globose, pale yellow-green, rugulose, 3 to 4m in diameter; perithecia 
enveloped by hiille cells, light brownish in color, spherical, partially em- 
bedded in the mycelial felt (fig. 42 D), about 125 to 150m in diameter includ- 
ing the enveloping hiille-cell layer, with perithecial wall 1-cell layer in 
thickness, ripening quickly and with ripe asci breaking down to leave the 
ascopores free; ascospores purple-red, lenticular, with smooth wall, with 
spore body 4 to 4.8m by 3.4 to 3.8m, and with two pleated equatorial crests 
about 0.5m in width paralleled by a secondary narrower pair (fig. 43 B) 
which are sometimes indistinct. 

Type NRRL No. 201 (Thorn No. 4138.N8) from New Jersey soil and kept 
in culture since 1916. Other strains examined include isolations from 
Texas, Colorado, Louisiana, and Maryland. 

Aspergillus rugulosus Thom and Raper, Mycologia 31: 660-663, 

fig. 4. 1939. 

Colonies on Czapek's solution agar slowly and restricted^ growing (fig. 
41 C), buckled or wrinkled in a mass 2 to 3 mm. deep, enveloping abundant 
perithecia at different depths, often eventually splitting in the central area, 
purple-gray to purple-brown in age, with green heads sparsely produced 
and hence not generally evident, occasionally seen as small groups and 
marginal extensions into drying media; reverse in shades of deep purple- 
red; conidial heads, short columnar, 75 to 100m by 30 to 40m; conidiophores 
sinuous, smooth-walled, pale brownish in color, 50 to 80m long, slender, 
varying up to 5m in width, then enlarging to vesicular hemispheres 8 to 10m 
in diameter; primary sterigmata 7 to 8m by 3 to 3.5m, secondary sterigmata 
6 to 7m by 2.5 to 3m; conidia globose, green, rugulose, 3 to 4m. 

Perithecia very abundant, often imbedded in the mycelium as 2 or 3 layers 
and each surrounded by hyphae and dark brown hiille cells (fig. 42 E), 
globose, 225 to 350m in diameter including mycelial coverings, with dark 
reddish-purple walls of one cell thickness, quickly ripening and breaking 
down to leave ascospores free; asci 10 to 11m in long axis; ascospores purple- 
red, lenticular, walls conspicuously rugulose (fig. 43 C), with spore bodies 4 



THE ASPERGILLUS NIDULANS GROUP 



161 



to 4A/J. by 3.6 to 3.8ju, and with 2 pleated equatorial crests with sinuate and 
entire margins about 0.5 to 0.6m in width. 



A : A. nidulans 



B : A. unguis 




immBm 




O A. nidulons vor. lotus 




D : A. quodrilmeotus 



IS&&>»S8J&&_ su k s&sg^^Ssfc^sisi&s 




E ■' A. rugulosus 



F : A. voriecolor 




Fig. 42. Diagrammatic representations of cross sections of colonies of different 
species of the Aspergillus nidulans group showing the relative abundance of conidial 
heads and perithecia and the manner in which these structures are borne : con, conidial 
heads; d.h., mantle of divergent hyphae; hul, huile cells; my, mycelial felt; per, 
perithecia; ps, pseudostalk of hiille cells and sterile hyphae; st, long, thick walled, 
sterile hyphae; sub, substratum; and un, unstalked perithecium. Scale approxi- 
mate. (Reprinted from Thom and Raper, "The Aspergillus nidulans Group," 
Mycologia 31: 653-669. 1939.) 



Cultures studied included Type NRRL No. 206 (Thom No. 4138.T11) 
from New Jersey soil, as discussed by Thom and Church in The Aspergilli, 
p. 138, and also isolates from Washington, D. C, Texas, Nebraska, and 
California. Very common in soil. 



162 



A MANUAL OF THE ASPERGILLI 




Fig 43 Ascospores of different species of the Aspergillus nidulans group. A, A. 
nidulans. B,A.quadrilineatus. C,A.rugulosus. D, A. vanecolor In each species 
upper left and right and center left spores represent surface profile views; center 
right, surface in face view; lower left, optical section in profile; and lower right, 
optical section in face view. (Reprinted from Thorn and Raper, 'The Aspergillus 
nidulans Group," Mycologia 31: 653-669. 1939.) 



THE ASPERGILLUS NIDULANS GROUP 163 

Culturally and microscopically the above strains present similar pictures, 
with the exception of the strain recently received from Bliss in California 
(NRRL No. 2 1 1 ) . In contrast to the others, this culture produces abundant 
conidial heads and relatively fewer perithecia. The ascospores and conidial 
structures, however, duplicate those of the typical strains; hence we do not 
at present feel warranted in designating this as a variety, or otherwise 
separating it from the species A. rugulosus. 

Aspergillus variecolor (Berk, and Br.) Thorn and Raper, in Mycologia 31: 

663-G67. fig. 4D and fig. 5. 1939. 

Synonyms : Emericella variecolor Berk, and Br. in Berkeley, Introd. 

Crypt, Bot. p. 340-341; fig. 76. 1857. See Patouillard, 

Bull. Soc. Myc. Fr. 7: 43-49. pi. 4. fig. 6-12. 1891. 
Inzengaea erythrospora Borzi, Jahrb. Wiss. Bot. (Pringsheim) 

16: 450-463. pi. 19, 20. (1884) 1885. 
Emericella medias Chowdhury and Mathur, Ann. Myc. 36: 

61-63. 1938. 
Aspergillus stellatus Curzi, Rend. Acad. Naz. Lincei 19: 

424-428. fig. 1. 1934. 

Colonies on Czapek's solution agar with vegetative mycelium largely 
submerged, sparse, spreading slowly in the agar, producing green heads 
freely in the center of the colony, less abundantly in the outer areas, large 
gray perithecia produced in clusters in colony center and at the margin in 
some strains, with smaller perithecia scattered through the intervening 
thinner areas of the colony (fig. 44 A), in other strains producing large 
perithecia abundantly throughout the colony (PI. IVD); reverse color in 
shades of purple-red. Conidial heads green, columnar (fig. 44 D), relatively 
long, mostly 100 to 200m, occasionally up to 300m by 30 to 40m; conidiophores 
arising directly from submerged hyphae, straight with smooth walls, cinna- 
mon-brown in color, mostly 140 to 200m long by 3 to 5m in diameter, 
broadening gradually to become hemispherical vesicles about 8 to 10m in 
diameter; primary sterigmata 7 to 8m by 3 to 4m, secondary sterigmata 8 to 
9m by 2.5 to 3m; conidia globose, rugulose, 3 to 3.5m; perithecia when clus- 
tered (fig. 44 Aa) 300 to 400m in diameter surrounded by a felt of hyphae 
and hulle cells and supported by masses of hyphae and hulle cells forming 
false stalks (fig. 42 F), giving the structures a pyriform appearance (fig. 
44 B) ; scattered perithecia much smaller (fig. 44 C) and with envelope of 
supporting cells often much reduced in mass; hulle cells abundant and 
essentially like those of the species A. nidvlans. 

Perithecial wall when stripped of enveloping cells purple-red, brittle, 
composed of a single layer of cells; asci quickly ripening and breaking down 
to leave the cavity filled with ascospores; ascospores purple-red, with spore 
bodies lenticular and 3.6 to 4m by 2.8 to 3m, with two prominent equatorial 



164 



A MANUAL OF THE ASPERGILL] 




Fig. 44. Aspergillus variecolor. A, Central area of colony; a, cluster of large 
pseudostalked perithelia; b, scattered, smaller, unstalked perithecia (see also figure 
42 F); c, scattered conidial heads, X 6. B, Enlarged view of large, pseudostalked 
perithecium, X 65. C, Enlarged view of small, unstalked perithecia, X 65. D, 
Enlarged view of conidial heads, X 65. (Reprinted from Thorn and Raper, "The 
Aspergillus nidulans Group," Mycologia 31: 653-669. 1939.) 



crests, up to 3.5m in width, pleated and cot to give a stellate appearance to 
the ascospores (fig. 43 D). 



THE ASPERGILLUS NIDULANS GROUP 165 

The above description is based primarily upon a culture received from 
Prof. Verona in Italy and carried in our collection as NRRL No. 212 (Thorn 
No. 5602.3). 

Bliss forwarded a culture (NRRL No. 214) isolated from date fruits in 
California which differs from the above in the following particulars: (1) 
Conidial heads are produced abundantly, (2) the mycelium is not predomi- 
nantly submerged, and (3) the colonies in reverse are deep purple. How- 
ever, the ascospores of the two strains are strikingly similar in size and 
pattern, the perithecia of each appear pyriform in shape, and the conidial 
structures of the two are essentially alike. More recently a strain has been 
isolated from Arizona soil (NRRL No. 1954) which produces abundant large 
"stalked" perithecia but very few small perithecia or conidial heads. 
There is, thus, evidence of considerable natural variation among members of 
this species. All of these strains have the same type of stellate ascospore. 

Since the type of ascospore described here had already been assigned to 
Emericella and Inzengaea, consideration of the literature of these genera is 
necessary. 

Emericella variecolor, genus and species new, was described by Berkeley 
and Broome in 1357 as doubtfully a Gasteromycete or possibly a lichen. 
The perithecium with a mass of stellate spores was considered as gastro- 
mycetous in character while what we now recognize as the "hulle" cells of 
Eidam (figured) suggested to them the possibility of an algal associate. 
Berkeley's material was also examined by Montague and part of it deposited 
in the Museum d'Histoire Naturelle de Paris. This was reexamined by 
Patouillard in 1891 and its ascomycetous nature determined. No conidial 
apparatus was found by either Berkeley or Patouillard. 

Inzengaea erythrospora as the type species of a new ascomycetous genus 
was figured and described by Borzi in 1885, showing stellate red ascospores, 
and the hulle cells of Eidam. Borzi 's figure showed a coremium-like conid- 
ial apparatus which was designated Coremium Borzianum by Saccardo. 
Ed. Fisher in 1893 transferred the species to Emericella of Berkeley and 
placed the genus next to Aspergillus in the "Pflanzenjamilien." Saccardo 
(in Syll. 9: 610) on the other hand accepted Inzengaea and dropped Emeri- 
cella because of the errors in description and placement by Berkeley. Borzi 
figured the spores of A. variecolor {Inzengaea erythrospora) correctly but 
obviously misinterpreted the germination of the ascospores since he showed 
them splitting as if turned 90°, bringing the crest perpendicular to the 
center of the valve instead of attached to its edges. 

There the taxonomic situation stood until Vuillemin in 1927 concluded 
that the ascosporic apparatus, the stellate red ascospores and the cells of 
Eidam, as clearly shown in their figures and material, showed the identity 



166 A MANUAL OF THE ASPERGILLI 

of Emericella and A. nidulans. He therefore transferred A. nidulans to 
Emericella as the oldest established genus and apparently did not even 
consider Inzengaea. 

Ciferri (1938) has recently completed a study of Emericella variecolor 
embracing cultural investigations together with a review of the literature 
of the genus. He did not recognize the close relationship of this fungus to 
Aspergillus nidulans, and was apparently unmindful of the likeness of their 
conidial structures and the essential similarity of their perithecia and asco- 

spores. 

In 1934 Curzi described as Aspergillus stellatus a fungus characterized by 
hulle cells and red, stellate ascospores. However, he apparently did not 
know of either Berkeley's or Borzi's earlier designation of a similar fungus. 
It is unfortunate that his exceedingly descriptive binomial must be reduced 
to synonymy. 

Fortunately for this discussion, cultures NRRL Nos. 212, 214, and 1954 
present both the stellate spores and apparently stalked perithecia figured 
by Berkeley, Patouillard, and Borzi. (Compare figures 42, F, and 44, 
B with Berkeley's figure 76a, Patouillard 's figures 7 and 8, plate 4, and Bor- 
zi's figure 10, plate 19.). This made possible a restudy of the whole 
morphologic situation from fresh material. The perithecial body itself 
was found not to be stalked but to rest upon a sterile mass of hulle cells 
and mycelium giving the superficial appearance noted by earlier observers. 

The coremium of Borzi remains unaccounted for. Obviously in Borzi's 
discussion the material was rotten olives and very old. No cultures were 
made. The conidia-producing apparatus (figured) differs essentially in 
type from the conidial apparatus of the Aspergillaceae with which Fischer 
correctly placed Emericella because of its perithecia and ascospores. We 
are convinced that the coremia belonged to some other fungus. 

It is not possible to separate the perithecium of this fungus from that of 
the other species in the A. nidulans group nor are there characters to take 
this type of perithecium out of a genus with the yellow perithecium of the 
great A . glaucus series of species which are widely known. Consistent with 
the policy of keeping the Aspergilli in one group, both Emericella and In- 
zengaea are dropped for purposes of this discussion. 

Aspergillus caespitosus Raper and Thorn, in Mycologia 36: 563-565, 

fig. 4. 1944. 

Colonies varying markedly upon different media; upon Czapek's solution 
agar rather slow growing, attaining a diameter of 6 to 8 cm. in three weeks 
at room temperature, plane or somewhat furrowed, mycelium largely sub- 
merged and extremely tough, tearing with difficulty, producing numerous 
dark green, hemispherical to loosely columnar heads in central colony areas, 



THE ASPERGILLUS NIDULANS GROUP 



167 



characterized particularly by clusters of irregular ovoid to elliptical, thick- 
walled hulle cells, at first colorless becoming reddish-purple in age, scattered 



**. 












8 









•- 






&>*5."» 













S85C 



• 











'•V* 



V 



"*»_-.-> 






Fig. 45. Aspergillus caespitosus ; , NRRL No. 1929. A, Portion of colony on 
Czapek's solution with 1 percent liver extract, showing crowded conidial heads in 
central portion and scattered hulle cell masses in surrounding areas, two weeks old, 
incubation at room temperature, X 1.8. B, Portion of colony enlarged showing 
hulle cell masses, X 10. C, Silhouettes of conidial heads developed on hay infusion 
agar, X 48. D, Typical conidial head showing form of vesicle and arrangement of 
sterigmata, X 600. E, Portion of htille cell mass showing irregular size and form 
of component elements, X 265. (Reprinted from Raper and Thorn, "New Species 
of Aspergilli from Soil," Mycologia 36: Nov.-Dec. 1944.) 

unevenly (fig. 45 A and B) or arranged in irregular concentric zones; reverse 
colorless at first, becoming dark reddish-purple in age, particularly beneath 
the hulle masses; odor none. Conidial heads dark dull yellow-green to 



168 A MANUAL OF THE ASPERGILLI 

empire green (Ridgway, PI. XXXII), generally hemispherical to loosely 
columnar, mostly 75 to 125m in diameter. Conidiophores straight or 
slightly sinuous (fig. 45 D), mostly 250 to 325m in length, occasionally up 
to 350m by 5.0 to 6.5m in diameter, of approximately uniform diameter 
throughout, relatively thick-walled, (1.2 to 1.5m in basal portion to 0.8 
to 1.0m in terminal area), smooth, tan to light brown in color. Vesicle 
slightly elongate, the upper hemisphere loosely covered by sterigmata 
(fig. 45 D), the lower half sterile and often lightly colored, mostly 15 to 20m 
in diameter. Sterigmata in two series (fig. 45 D), primaries normally 6.5 
to 8.5m by 3.5 to 5.0m, secondaries 6.5 to 8.0m by 3.0 to 4.5m, typically bottle 
form but commonly much swollen and often quite irregular in form and 
dimensions. Conidia globose, spinulose, green, mostly 3.5 to 4.5m, rarely 
larger, hiiHe cells very abundant, thick-walled, irregularly globose, ovoid or 
elliptical (fig. 45 E), ranging from 12 to 18m in globose cells to 12 to 15m 
by 25 to 30m in the most elongate bodies, forming compacted masses of 
indefinite size, extremely tough and in age becoming almost sclerotioid, at 
first colorless but in age characterized by an abundant reddish-purple 
intercellular pigmentation. 

Colonies upon malt agar characterized by a dense stand of erect conidio- 
phores bearing hemispherical to radiate or loosely columnar heads of dark 
green color approximately empire green (Ridgway, PL XXXII) and the 
complete absence of hulle cells; reverse in light brown shades; odor none. 
Details of morphology as upon Czapek's solution agar. 

Colonies upon hay infusion agar like those upon malt except less heavily 
sporing. 

Strains include NRRL No. 1929 (type) isolated from Arkansas soil and 
other isolations from Arizona and Texas soils. 

This species is of particular interest because of its apparent transitional 
position between the A. nidulans group and A. ustus. In the character 
of its conidiophores, its reddish-purple pigmentation, and in the general 
color and markings of its conidia it retains the characters of A. nidulans 
and closely related species. In the absence of fertile perithecia and asco- 
spores, the predominantly hemispherical shape of its conidial heads, and in 
the variable and irregular form of its hiiHe cells, it is strongly suggestive of 
the A. ustus series. While we are convinced of its. intermediate position 
between the A. nidulans group and A. ustus, we place it with the former 
since we believe it is most closely allied to this group. It is believed sig- 
nificant that superficially, cultures of Aspergillus caespitosus and Aspergillus 
variecolor (Berk, and Br.) Thorn and Raper (1939) are strikingly similar 
upon Czapek's solution agar. This similarity is particularly marked when 
plates are viewed in reverse since an intense pigmentation marks the under 
surface of perithecia in the latter case and the under surface of older hulle 
masses in the former. 



THE ASPERGILLUS NIDULANS GROUP 



169 



Aspergillus unguis (Emile-Weil and Gaudin) Emend. Thorn and Raper, 

Myc. 31, p. 667, fig. 6. 1939. 

Synonyms: Sterigmatocystis unguis Emile-Weil and Gaudin, Arch. Med. 
Expt. Anal. Path. Paris 28: 463-465, fig. 4, 1919. 
A. loakiashanensis Shih, Lingnan Sci. Jour. 15 (3): 369. p. 
16, fig. 2. 1936. 

Colonies on Czapek's solution agar restrictedly growing, plane, spreading 
at the margin as irregular lobes (fig. 41 E), yellowish-green, green to dark 
green becoming brown in age; without perithecia or hulle cells. Mycelial 
preparations show striking sterile, thick-walled hyphae with walls in brown 




Fig. -46. Sterile spicule hyphae of Aspergillus unguis. A, Cluster of sterile 
hyphae, X 370. B, Apex of sterile hypha, X 740. C and D, Mid-portions of sterile 
hyphae showing thick roughened walls, X 740. (Reprinted from Thorn and Raper, 
"The Aspergillus nidulans Group," Alycologia 31: 653-669. 1939.) 



shades, irregularly roughened (fig. 46), tapering to a blunt point, arising 
sometimes from foot-cells suggesting the origin of conidiophores, sometimes 
apparently from mycelial cells, often up to 1,000m or more in length, slant- 
ing upward but usually rising only slightly above the conidial area (fig. 
42 B). 

Conidial heads columnar, 75 to 150m by 40 to 50m; conidiophores smooth- 
walled, dull brown in color, mostly 45 to 65m in length by 3 to 5m in diameter, 
enlarging to vesicular hemispheres 9 to 12m in diameter; primary sterigmata 
5 to 6m by 2.5 to 3m, secondary sterigmata 5 to 6m by 2 to 2.5m; conidia 
globose, rugulose, dull green, 2.5 to 3.5m in diameter. 

Cultures of A. unguis are obtained frequently from medical laboratoies 
apparently as more or less active pathogens but occasionally isolated from 



170 A MANUAL OF THE ASPERGILLI 

soil and decaying organic matter. The question whether the non- 
ascosporic members of the group have merely dropped the ascogenous 
phase or constitute a separate species was answered when more complete 
examination showed the sterile or spicule hyphae to be regularly produced 
in the non-ascosporic, but never found in ascosporic series. 

Pathogenicity 

Aspergillus nidulans in some of its forms and variants has been demon- 
strated as a parasite in human nails (onychomycosis), often enough to 
establish its pathogenicity. A. nantae Pinoy (1927) probably belongs 
here although the data are mainly pathological, hence not adequate for 
definite identification of the organism. A. nidulans forme cesarii Pinoy 
(1915) isolated from a mycetoma of the lung of a donkey, and A. nidulans 
var. nicollei Pinoy (1906) isolated in Tunis from a case of mycetoma, or 
madura foot, represent additional strains which were at least secondary 
pathogens. The nearly related A. unguis is usually the more common 
form isolated from human material. A. Brodeni (Mattlet) Dodge (1935) 
from a bronchomycosis in Africa might have been close to A. unguis. A. 
nidulans and A. unguis are both widely distributed as saprophytes; hence 
are constantly encountered as components of dirt reaching the extremities 
by contamination. Infection of the air passages is comparatively rare. 

Occurrence and Economic Importance 

In addition to their role as occasional disease producing agents, members 
of the A. nidulans group are believed to be significant in decomposition 
processes. They are among the molds most commonly isolated from soil, 
and very frequently appear in considerable abundance upon vegetable ma- 
terial undergoing slow decomposition. Aspergillus rugulosus, A. quadriline- 
atus, and A . caespitosus occur most frequently in soils from the comparatively 
dry, warm soil of Texas, Arizona, and adjoining areas. Aspergillus varie- 
color has been isolated from olives in Italy, date fruit in California, and from 
Arizona soil. Aspergillus nidulans is abundant and cosmopolitan in its 
distribution. 



Chapter XII 
THE ASPERGILLUS USTUS GROUP 

Outstanding Characters 

Colonies more or less floccose, at first white but becoming dull in age, in 
most members varying from olive-gray through reddish-brown to 
fuscous, as conidial structures develop. 

Conidiophores in yellow-brown shades; smooth. 

Heads irregular in form, ranging from more or less radiate to hemi- 
spherical to loosely columnar. 

Vesicles hemispherical; sterigmata in two series, loosely arranged. 

Conidia roughened, 3.0 to 5.0m, varying from echinulate to marked with 
conspicuous color bars, and ranging in color from pale blue-green 
through olive-green shades to deep brown (fuligineus). 

Hulle cells regularly present, thick-walled, elongate, often more or less 
curved and twisted. 

Included here are representatives of a most abundant and widespread 
group of fungi, especially common in soil and upon decaying vegetation. 

Group Key 

Colonies predominantly floccose, heavy sporing, hiille cells not aggregated in small 

clusters A. ustus (Bain.) Thorn and Church 

Colonies more or less floccose, light sporing, hiille cells aggregated in small clusters 

A. granulosus Raper and Thorn 

Aspergillus ustus (Bainier) Thorn and Church, in The 
Aspergilli, p. 152. 1926. 

Synonym : Sterigmatocystis usta Bainier, in Bui. Soc. Bot. France 28 : 
78. 1881. 

Colonies upon Czapek's solution agar spreading broadly, plane, sulcate, 
or umbonate, rarely zonate, more or less felted or floccose; at first white, 
becoming olive-gray, yellow-brown, fuscous or russet to purplish vinaceous 
with the development of mature conidial structures (PI. IV E, and figs. 
48 A and B) ; generally heavy sporing, with some conidiophores arising from 
the substratum but more abundantly from aerial hyphae ; reverse in shades 
of yellow, orange, and brown to almost black in age; odor not pronounced. 
Heads radiate to irregularly hemispherical, sometimes loosely columnar, 
commonly splitting into more or less well-defined columns in age, very 
variable in size, ranging in color from dull green or olive-gray, through gray- 

171 



172 



A MANUAL OF THE ASPERGILLI 



ish-brown to fuscous or fuligineus. Conidiophores arising from submerged 
hyphae (fig. 47 A) ranging up to 500m long by 3 to 6m, aerially borne conidio- 
phores, ranging from very short (fig. 47 Ai) up to 125m by 2 to 5m, sinuous, 
sparsely septate, with walls rather thin, smooth, and uniformly colored 
some shade of brown. Vesicles hemispherical to subglobose, 8 to 20m in 
diameter, smaller in some strains (fig. 47 A). Sterigmata colorless or 










Fig. 47. Aspergillus ustus group, X 840. A, Typical conidial head showing com- 
paratively loose sterigmata in two series and conspicuously roughened conidia, strain 
NRRL No. 278. A u Diminutive head, as often seen in strain NRRL No. 275. B, 
Typical head of Aspergillus granulosus, NRRL No. 1932. 



colored, semi-radiate, loosely arranged into two series, primary sterigmata 
4 to 7m by 3m, secondary sterigmata 5 to 7m by 2.0 to 2.5m- Conidia globose, 
3.5 to 5.0m, roughened, echinulate to marked with conspicuous color bars, 
ranging from greenish through olive-gray to yellow-brown or fuligineus. 
Many strains producing thick-walled hulle cells (fig. 49 E) ranging in form 
from irregularly ovate or elongate in some strains, to serpentine, helicoid, 
or twisted in others, essentially as in Aspergillus flavipes. 



THE ASPERGILLUS USTUS GROUP 



173 



Upon malt agar, colonies frequentl,y heavier sporing and conidial heads 
generally tending to run to dull gray-green shades rather than brown. 

Very common in soil and decaying vegetation. Species diagnosis repre- 
sents a composite based upon many isolations from this country and abroad. 





B 




D 



Fig. 48. Aspergillus ustus: cultures growing upon Czapek's solution agar at room 
temperature, 10 days. A, Strain XRRL No. 275 characterized by loose floccose 
colonies and moderate sporulation. B, Strain NRRL No. 278 characterized by 
heavier spore production and the presence of abundant hiille cells. C, A. ustus var. 
laevis, NRRL No. 1852, characterized by loose floccose colonies and conidial heads 
often near brick red in color. D, Strain NRRL No. 1974 characterized by the pro- 
duction of very abundant hiille cells in concentric zones. 



Individual strains differ markedly in their general habit and colony colora- 
tion, in the color of their fruiting structures, in the marking and coloration 
of their conidia, in the presence or absence of hiille cells, and in the form of 
these structures when present. By a deliberate selection of strains, one 
can find sufficient difference to warrant the assignment of specific designa- 



174 A MANUAL OF THE ASPERGILLI 

tions to particular cultures. Yet all possess the characters noted above and 
so constitute a well-defined group whose variations are matters of detail. 
Such differences as occur, moreover, tend to become bridged as comparative 
cultural and microscopic studies of many isolations are made ; hence the use 
of these differences becomes of questionable value for diagnostic purpose. 
We have considered it desirable, therefore, to include the whole series under 
one name as a single species aggregate, Aspergillus ustus, and to call atten- 
tion to some of the major differences which one may expect to encounter 
among the members of this species. Although Bainier was not sufficiently 
explicit in his description of Sterigmatocystis usta to enable us to identify 
with certainty the form with which he worked, his usage is accepted upon 
(1) the basis of priority, and (2) the receipt of a culture (Thorn No. 4640. 
488) from his laboratory labeled Sterigmatocystis usta, which possessed the 
basic characters of the group as herewith set forth. 

Long after the publication of Sterigmatocystis usta, Bainier described a 
second species belonging to this group, Sterigmatocystis insueta (Bui. Soc. 
Mycol. France 24: 85-87, PL VIII, Fig. 1-13. 1908), and emphasized 
the fuligineus character of its colonies, the predominant origin of brown 
fruiting structures from aerial mycelium, the larger size and the darker 
color of its conidia, which were characterized by the presence of pronounced 
color bars. Strains showing these characters probably represent the type 
most commonly encountered among miscellaneous isolations of forms be- 
longing to this group. Recognition of these forms as constituting a distinct 
species has been considered, but in the absence of any clearly definable line 
of separation from A. ustus it is believed desirable to leave them within the 
somewhat extended framework of this species. Cultures possessing these 
characteristics very commonly exhibit hiille cells varying in different strains 
from ovoid to irregularly elongate, to serpentine, helicoid or otherwise 
twisted. One strain showing much-twisted hiille cells was isolated and 
contributed by Thaxter under the manuscript name A. helicophorus. 
Typically, the conidiophores of these forms are grayish-brown, commonly 
quite dark. The vesicle and sterigmata are likewise frequently colored. 
Conidial color as seen under high magnifications varies with age from pale 
to olive green to fuligineus, and conidial markings from fairly coarse 
echinulations to intensively colored bars and tubercles. 

Possibly unaware of the existence of Sterigmatocystis usta and S. insueta 
(since no mention is made of either species), Abbott (1926) subsequently 
described Aspergillus minutus. His description indicated that he was 
dealing with a strain essentially similar to those considered by Bainier as 
S. insueta. This is confirmed by examination of his type culture, NRRL 
No. 283 (Thorn No. 4894.2). A second culture, NRRL No. 285 (Thorn 
No. 4894.1), received from Abbott under the manuscript name A. humus 
likewise represents a member of this series and differs from the more 



THE ASPERGILLUS USTUS GROUP 175 

common forms only in its more floccose habits and in the production of 
somewhat smaller conidial heads. 

Among the more striking members of the Aspergillus ustus series 
examined are two isolations from soil collected in Panama and Mexico, 
respectively, which upon Czapek's solution agar normally produce strongly 
zonate colonies consisting of alternating areas of crowded conidial heads 
and heavy hiiHe cell development (fig. 48 D). Except for this difference in 
colony appearance, however, these strains appear to be typical of the species 
as described above. 

A single strain, NRRL No. 1852, isolated from Louisiana soil, possesses 
conidial heads of a dull brick red color (Ridgway, PI. XXXIX: russet 
vinaceous to sorghum brown) and abundant, much-twisted hulle cells. 
While Blochwitz's description is too inadequate to permit of detailed com- 
parison, it is suggested that this strain may represent his Aspergillus ustus 
var. laevis (Ann. Mycol. 32(1/2): 4. 1934), which was described as 
characterized by "red conidia and crooked hiiHe cells." In strain NRRL 
No. 1852, the conidia are conspicuously reddish en masse but appear only 
slightly colored when viewed with high magnifications. In contrast to 
most strains of the Aspergillus ustus group, the conidia are finely echinulate 
rather than coarsely roughened. It is suggested that this form (fig. 48 C) 
may represent a transition in the direction of Aspergillus flavipes since the 
latter species is likewise characterized by much-twisted hiiHe cells, brown 
conidiophore walls, and conidia which may show a reddish color in some 
strains but are smooth in all. 

Aspergillus granulosus Raper and Thorn, in Mycologia 36: 565-568, 

fig. 4. 1944. 

Colonies upon Czapek's solution agar growing well, attaining a diameter 
of 8 to 10 cm. in two to three weeks at room temperature, plane or irregu- 
larly furrowed, predominantly floccose, uneven in texture, buff to dull brown 
in color from felted sterile mycelia ; conidial heads few in number and gen- 
erally arising from the substratum direct, less often from aerial hyphae, 
commonly appearing in clusters, pale blue-green in color; colonies charac- 
terized particularly by abundant small, colorless clusters of irregularly 
globose, ovoid, or elliptical thick-walled hiiHe cells which superficially sug- 
gest perithecial initials and which in mass give to the colony a semi-granular 
appearance (fig. 50 B, E, and F); reverse in shades of dull yellow and 
brown; slight mushroom odor. Conidial heads few in number, commonly 
clustered in small groups, most abundant at colony margin, sometimes 
occurring on tufts of aerial hyphae, hemispherical to radiate, 75 to 125/x 
in diameter, very loose, consisting of comparatively few divergent spore 
chains (fig. 50 C), approximately pale niagara green in color (Ridgway, 
PI. XXXIII). Conidiophores erect, straight, nonseptate, mostly 350 to 



/* 



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Fig. 49. Htille cells. A, Characteristic thick-walled, globose to subglobose hiille 
cells of the Aspergillus nidulans group, A. variecolor, NRRL. No. 1954, X 450. B, 
Aspergillus caespitosus, strain NRRL No. 1930, hiille cells irregular in form and 
often poorly developed but of the same general pattern as A. nidulans, X 275. C, 
Aspergillus janus, NRRL No. 1787, hiille cells approximately globose and of the 
same basic type as in A. nidulans, X 450. D, Aspergillus granulosus, NRRL No. 
1932, hiille cells globose to ovoid, somewhat intermediate between A. nidulans and 
A. ustus, X 275. E, Aspergillus ustus, NRRL No. 280, characteristic hiille cells, 
elongate and much twisted, X 450. Hiille cells in Aspergillus flnvipes are of the 
same pattern. F, Aspergillus carneus, NRRL No. 1926, heavy walled, strongly 
septate mycelium suggestive of hiille cells, X 275. 

176 



THE ASPERGILLUS USTUS GKOUP 



177 



500^ in length, 5.5 to 8.0/x in diameter, approximately uniform in width 
throughout, thin-walled, smooth, tan to light brown in color, often slightly 




D 



Fig. 50. Aspergillus granulosus; NRRL No. 1932. A, Portion of colony on hay 
infusion agar showing clusters of conidial structures and small masses of hiille cells, 
two weeks old, incubation at room temperature, X 1.5. B, Portion of marginal area 
of two colonies on malt extract agar showing characteristic granular appearance 
resulting from numerous small clusters of hiille cells, X 1.5. C, Silhouettes 
of conidial heads, X 48. D, Conidial head showing slightly elongate vesicle, sterig- 
mata in two series, and constriction in conidiophore just beneath the vesicle, X 750. 
E, Small clusters, or "granules", of hiille cells, X 48. F, Double cluster of hiille 
cells, much enlarged, X 265. (Reprinted from Raper and Thorn, "New Species of 
Aspergilli from Soil," Mycologia 36: Nov .-Dec. 1944.) 



constricted just beneath the vesicle (figs. 47 B and 50 D). Vesicle ovate 
to elliptical, thin-walled and easily broken, largely covered by sterigmata, 
12 to 18^ m diameter by 15 to 25/z in length. Sterigmata in two series, 



178 A MANUAL OF THE ASPERGILLI 

both comparatively short and stout; primaries 3.5 to 5.0m by 3.0 to 4.0m; 
secondaries 4.0 to 5.5m by 3.0 to 3.5/z, commonly bottle-form. Conidia 
globose, pale green, delicately echinulate. Mostly 4.8 to 5.5/x in diameter, 
rarely larger. Hiille cells abundant, irregularly globose, ovoid or somewhat 
elongate, commonly 12 to 30m in long axis, walls heavy, 4 to 5m in thickness, 
borne primarily in small, colorless clusters which are quite conspicuous at 
colony margins and lend to them a characteristic granular appearance. 

Colonies upon malt extract agar showing an accentuation of hiille cell 
development (fig. 50 B) and a reduction in conidial heads. Otherwise 
duplicating the cultural picture presented upon Czapek's solution agar. 

Colonies upon hay infusion agar (fig. 50 A) thin but broadly spreading, 
characterized by scattered clusters of hiille cells and erect conidial fructifica- 
tions giving to the culture a sparsely granular appearance. 

Type culture NRRL No. 1932 was isolated in 1942 from a sample of soil 
collected in Fayetteville, Arkansas, and contributed by Mr. F. R. Earle. 
Additional strains have been isolated from soils collected in Texas, Arizona, 
and Costa Rica. It is believed common in soils where the temperature 
remains at a high level during part or all of the year. 

Different strains vary materially in the number of conidial heads pro- 
duced upon common laboratory media such as Czapek's solution and malt 
extract agars, ranging from abundant heads in some to only widely scattered 
heads in others. All fruit reasonably well, however, upon hay infusion agar, 
the medium upon which original isolations were made. 

The brown color of the conidiophores, the presence of ovoid to somewhat 
irregular hiille cells, and the green color of its conidia place this species in 
the group with A. ustus. It differs markedly from the more common 
representatives of this group, however, in the lighter and persistently green 
color of its conidia, the small clusters rather than irregular masses of hiille 
cells, and in possessing somewhat more elongate vesicles. In this latter 
character it suggests Aspergillus flavipes but is in turn excluded from this 
group by the green color of its spores. 

Occurrence and Economic Importance 

Representatives of the Aspergillus ustus group are perhaps the most 
abundant of all aspergilli in soil. They regularly occur in large numbers 
and in considerable variety. The common species, A. ustus, occurs alike 
in cultivated and forest soils and in approximately equal abundance in 
soils from southern and from north temperate areas. A. granulosus, on 
the contrary, has been isolated only from southern sources. Members of 
the group are not known to be particularly active agents of decomposition, 
but their great abundance in nature is believed indicative of a significant 
role in many decay processes. 

Their biochemical activities and potentialities are almost completely 
unknown. 



Chapter XIII 
THE ASPERGILLUS FLAVIPES GROUP 

Outstanding Characters 

Conidiophores smooth, in some shade of yellow, with color often confined 

to the outer layer. 
Heads barrel-form to columnar when well developed, white or slowly 

becoming some shade of vinaceous-buff to avellaneous (Ridgway, 

PL XL). 
Vesicles subglobose to elliptical. 
Conidia colorless, smooth, thin-walled. 
HiiHe cells generally present, helicoid or variously twisted. 

The range of variation presented within the group has led workers with 
only a few representatives before them to offer names and descriptions for 
the strains under observation. However, when large numbers of strains 
are brought together and cultivated upon a considerable range of substrata 
the continuity of the group as a natural and related series of strains becomes 
apparent. Further study of this whole group may lead to separation upon 
lines accounting for certain published descriptions not at present identifi- 
able. For purposes of the present manual, however, it is believed desirable 
to broaden the description of Aspergillus flavipes (Bain, and Sart.) Thorn 
and Church sufficiently to include a closely related series of organisms 
rather than restrict it to the particular strains studied by Bainier. 

Aspergillus flavipes (Bain, and Sart.) Thorn and Church, 
The Aspergilli, p. 155. 1926. 

Synonym: S. flavipes Bainier and Sartory (Bui. Soc. Myc. France 27: 
90-96, PL HI., fig. 1-6. 1911). 

Colonies upon Czapek's solution agar rather slow growing, becoming 3 
to 5 cm. in diameter in about 10 days; mycelium yellowish, dull buff, com- 
monly becoming brownish in age; heads pale to dull buff in some strains 
to avellaneous or even very light cinnamon in others (PL IV F and fig. 
51 A), submerged mycelium persistently colorless in some strains, develop- 
ing many shades of yellow, orange, orange-brown to red (Madder-brown 
of Ridgway, PL XIII) or almost black in reverse of colonies in others; 
in some strains producing at the surface of the agar closely woven yellow 
to orange masses of hyphae enmeshing numerous helicoid or variously 
twisted, thick-walled hiille cells; occasional strains showing dark masses 

179 



ISO 



A MANUAL OF THE ASPERGILLI 



suggestive of sclerotia; aerial mycelium more or less abundant, colorless 
or yellow to orange; producing in many strains numerous large drops of 
transpired fluid, pale to yellow or orange-red (acting as an indicator, chang- 
ing from yellow with acid to orange-red with alkali) ; commonly 




Fig. 51. Aspergillus Jlavipes group. A, 
Czapek's solution agar. 2 weeks, room tei 



.4. flaripes XRRL No. 295 growing on 
temperature. B, Strain XRRL No. 287 of 
the same species, found bv White (1943) to produce a penicillin-like substance. C, 
Strain XRRL Xo. 1959 characterized by the production of an excessive amount of 
exudate. D, Photomicrograph of a typical head showing the elongate vesicle and 
crowded sterigmata in two series, X "50. 



characterized by a disagreeable odor approaching putridity. Heads 
typically becoming columnar masses, shading from persistently white, 
through shades of pale to deep avellaneous as in A. terreus; but usually in 
rather sharp contrast to the color of the mycelium. Conidiophores from 
300 to 500m by 4 to op. in crowded areas, up to 2 to 3 mm. in length and 



THE ASPERGILLUS FLAVIPES GROUP 181 

8 to 10/x in diameter in some strains and under some conditions of culture, 
walls more or less yellow under the microscope, with color mostly localized in 
the outer layers of the cell-wall and occasionally as disk-like concretions 
on the surface of walls otherwise smooth. Vesicles subglobose to elliptical 
(fig. 51 D), up to 30 by 4G> in the largest forms, usually with diameter 
twice that of the conidiophore in smaller forms. Sterigmata in two series, 
colorless or nearly so, closely packed over the apex of the vesicle in small 
heads, and covering the vesicle in large heads, primary sterigmata about 
6 or 8m by 2 to 3/x, secondary sterigmata 5 to 8/x by 1.5 to 2/x. Conidia 2 
to 3ju, smooth, subglobose, colorless or nearly so under high magnification, 
with chains aggregated to form columns as seen with the handlens in old 
cultures. 

Cosmopolitan in distribution and particularly common in soil and upon 
decomposing organic materials. 

Historically, the name applied to the series is taken from strains 4640.474 
and 4640.402 (Thorn Collection) obtained through daFonseca from the 
Bainier collection in Paris in 1922. These strains showed smooth yellow 
con idiophores 300 to 400m by 3 to 4ju, contrasting with heads that were 
rath er persistently white and possessed the general morphology of the series 
as described above. Hulle cells were not found. Nevertheless, the close 
relationship of these organisms to a great series of cultures obtained from 
many sources in which these structures are regularly found justifies us in 
broadening the use of the name. 

Culture No. 4640.486 (Thorn) received from the Bainier collection as 
S. rubescens (XRRL No. 291) shows deep floccose colonies with few heads 
and scattered dark Ivyphal masses in age. Hulle cells have not been seen in 
this strain. 

Among forms commonly obtained from soils collected from widely 
scattered areas in this country and abroad, a series of isolates seems to 
comply with the description given by Blochwitz for Aspergillus archi- 
flavipes (Ann. Mycol. 32(1/2): 84. 1934). This species as described 
represents an extreme development toward radiate heads, abundant conidia, 
conidiophores 2 to 3 mm. in length and the development of deep brown or 
actual red shades of color in the mycelium. Several strains observed in 
culture approach this description (fig. 51 C). Heads are at first globose, 
then become slowly barrel-form, i.e., short stocky columns. Large brown 
drops of transpired fluid are commonly seen which become yellow when 
acidified and return to reddish shades when alkali is added. Recognition 
of the species does not appear warranted since no clearly definable character 
exists which distinguishes these isolates from the less colored forms generally 
considered as representing Aspergillus flavipes in a more restricted sense. 



182 A MANUAL OF THE ASPERGILLI 

Antibiosis 

Working with a culture of Aspergillus flavipes from Thorn (No. 4303.46), 
NRRL No. 287, White (1943) has recently demonstrated the production of 
an antibacterial substance which in its action against Staphylococcus aureus 
strongly resembles penicillin. The substance was produced in greatest 
amount in a medium containing 5 to 10 percent corn steeping liquor as the 
sole nutrient. The addition of sugar is reported as definitely deleterious. 

Occurrence and Economic Importance 

Members of the Aspergillus flavipes group like hose of the A. ustus, 
versicolor, and terreus groups are cosmopolitan in distribution, and are 
especially common in fertile soil and upon decaying vegetation. They are 
not known to be active agents of decomposition but are capable of growing 
in the presence of a limited amount of water, hence are probably significant 
in initiating or continuing processes of decay where most micro-organisms 
are incapable of growing. Little is known of the biochemical activities or 
products of these forms. 



Chapter XIV 
THE ASPERGILLUS VERSICOLOR GROUP 

Outstanding Characters 

Conidial heads hemispherical to almost globose, in many different shades 

but usually showing green or blue-green. 
Conidiophores smooth, colorless, more or less sinuous. 
Vesicles globose to ovate or elliptical with radiate sterigmata borne over 

the upper half to three-fourths of the surface. 
Sterigmata in two series. 
Spores globose or subglobose, echinulate. 
HiiHe cells found in occasional strains, globose. 

Members of the Aspergillus versicolor group are cosmopolitan in distribu- 
tion. They occur regularly in soil, upon decaying vegetation, upon stored 
grains, upon cured meats, and upon a multitude of other products exposed 
to occasional moist air or undergoing slow decomposition. The morphology 
of all strains (with the exception of Aspergillus j anus) is basically alike, but 
different strains vary tremendously in their cultural appearance. This is 
especially true of Aspergillus versicolor where conidial heads in different 
strains vary in color from dark blue-green through green to yellow-green, 
to yellow and orange, and finally in some strains to yellowish-cream, buff or 
flesh color. 

Group Key 

I. Heads typically globose, less commonly hemispherical; blue-green in color with 
the blue element dominant; colony reverse and substratum usually in red or 
maroon shades A . sydowi series 

A. Conidial heads always blue-green, globose to radiate 

A. sydowi (Bain, and Sart.) Thorn and Church 

B. Conidial heads of two types: (a) blue-green in color, vesicles subglobose to 

elongate, borne on short conidiophores, and, (b) white, clavate, borne 
upon long conidiophores A. janus Raper and Thom 

II. Heads hemispherical or nearly globose at times; in green shades without blue 
admixture, buff to orange-yellow or occasionally flesh colored; colony reverse 
usually in pink, yellow-red or purple-red shades, rarely almost colorless 

A. versicolor series 

A. Conidia echinulate A. versicolor (Vuill.) Tiraboschi 

B. Conidia smooth A. humicola Chad, and Sach. 

183 



184 



A MANUAL OF THE ASPERGILLI 



Aspergillus sydowi (Bain, and Sart.) Thom and Church, The Aspergilli, 

p. 147. 1926. 

Synonym: S. sydowi Bainier and Sartory, in Ann. Mycol. 11: 25-29, PI. 
Ill, 1913. Compare PI. IV, cultures No. 4235.17 and K22, 
Thom and Church, The Aspergilli. 192(5. 

Colonies upon Czapek's solution agar growing well at room temperature, 
in most strains close-textured and velvety from crowded conidiophores and 
heads arising from the substratum (fig. 53 A), in other strains more or less 











■ *\ 

■ 

- 

1 


i # « * 


o 











1 ~"' r* 




•■* H 


\ 
A 





Fig. 52. Conidial structures in Aspergillus sydowi, NRRL No. 250, X 800. A, 
Typical, large conidial head borne upon an erect conidiophore arising from the 
substratum. B, Small head, borne as a lateral branch on an aerial hypha, consisting 
of a small cluster of double sterigmata. C, Minute heads in which sterigmata appear 
in a single series only. 

floccose (fig. 53 B) from interlacing and trailing aerial hyphae bearing con- 
idial heads ranging from large, well-formed structures to minute fruits 
consisting of clusters of simple sterigmata bearing few conidial chains, blue- 
green in color, approximately Delft blue or deep Delft blue (Ridgway, PI. 
XLII) with the blue effect especially marked in young fruiting areas (PL 
V A), reverse usually in shades of red, from coral red to maroon (Ridgway, 



Plate V 

A^feS »S?&* , aSa CB &S SK^iS "^ C , hUr ? h ^ R . RL N °- 25 °- * C«PP- right) 
1'iraboschi, NRRL No 239 D fcen e'r riJhn j.„ u ^ fcente-i- 1-ef t ) , AapnyiUiu versicolor (Vuill 
Clower left), Aspergillus te w ,fi NRRL v?^ "1t»'' S "'° °'' tVll, , 1U T / rab ^chi, NRRL No. 233. £ 
Tins objective could not be attainedTn making theit iS "^ ""gn^uW be in cinnamon shades, 
(v. Tiegh.) Blochwitz, VrSSO^^^^^^^T^ aJPT* ^^ f**"**" """"" 
were encountered in reproducing the colore i 7th stZ ) K.,™ S u S vinaceous fawn. Difficulties 
all other cultures growing upon CWk's sol ht inn »i« r> i f ?' cult " re ««>wing on malt extract agar; 
Research Laboratory. Kduc£ P ^ Northern Regional 



THE ASPERGILLUS VERSICOLOR GROUP 



185 



Pis. XIII and I) to almost black in some strains in age. Conidial heads 
typically radiate to nearly globose (fig. 52 A), ranging from 100 to 150m, 
but often reduced to small penicillate clusters of sterigmata especially in 
marginal colony areas and upon aerial hyphae (fig. 52 B and C). Conidio- 




Fig. 53. Colony types in the Aspergillus versicolor group, on Czapek's solution 
agar, 10 days, room temperature. A, Typical heavy sporing strain of A. sydowi. 
B, Floccose, lighter sporing strain of the same species. C,A. versicolor, XRRLXo. 
239, characterized by abundant green conidial heads and the production of deep 
red exudate. D, A. versicolor, XRRL Xo. 227, characterized by flesh-colored conidial 
heads. 



phores mostly arising from submerged hyphae, up to 500m in length by 5 
to 8m in diameter, colorless, smooth, comparatively thick-walled. Vesicles 
nearly globose, fertile over almost the entire surface, up to 20m in diameter. 
Sterigmata in two series; primary 6 to l\x by 2 to 3m, secondary 7 to 10m by 
2 to 2.5m- Conidia globose 2.5 to 3.0m (Bainier), in our culture up to 3.5m 
in diameter, conspicuously spinulose, green en masse. Globose hulle cells 



186 A MANUAL OF THE ASPERGILLI 

closely resembling those of the A. nidulans group have been seen in occa- 
sional strains. No sclerotia or perithecia are found, but clamydospores in 
solid substrata are reported. 

A typical member of our series of isolations was sent to Bainier who con- 
curred in our interpretation of his species. 

Reduced conidial apparatus appears in varying degree in all strains of 
A. sydowi examined. In typical strains, primary sterigmata, with their 
clusters of secondaries each bearing a chain of conidia, are found singly 
or variously grouped along trailing aerial hyphae. In other strains there is 
a progressive development of aerial mycelium, in the form of trailing hyphae 
either single or in ropes, coupled with a reduction in the number of typical 
A. sydowi heads. Strains are even occasionally seen in which only a few 
A. sydowi conidiophores and heads are found in what is otherwise a penicil- 
lium-like colony. Thus a series of strains exists which shows a fairly com- 
plete gradation from Bainier and Sartory's Sterigmatocystis sydowi to 
Penicillium restrictum of Gilman and Abbott. We are led to believe that 
the latter species should probably be assigned to this section of the genus 
Aspergillus. 

While morphologically very close to A. versicolor, A. sydowi is easily pre- 
sumptively recognized by the characteristic blue-green color of its conidial 
heads and the red colors in the substratum. A partial list of the sources of 
the isolations studied includes soil in Washington, D. C, Illinois, Manitoba, 
Florida, and Ceylon; moldy silk from a stocking factory; concentrated sugar 
products from Louisiana; dried fish in Japan; and bee-hives in Michigan. 
It is world-wide in distribution and very adaptable to substrata of widely 
different nature. 

Probable Synonyms 

Aspergillus tiraboschii Carbone (Atti d. Inst. Bol. Univ. Pavia Ser. II. Vol. XIV, 
p. 320, 1914) is described in the colors of A. versicolor but with the head of A. sydowi. 
It would appear to be more or less intermediate but, unless reisolated, must be 
dropped because of incomplete data. 

Sterigmatocystis tunetana Langeron (Bull. Soc. Path. Exot. 17: 345-347, text fig., 
1924) . This mold was recorded as isolated from an ulcer of the hand but failed to 
produce lesions in animal tests; as described the colonies were blue-green as in 
A. sydowi. 

A. sydowi var. achlamydosporus Nakazawa, Simo and Watanabe (Jour. Agr. Chem. 
Soc. Japan 10(2): 178-179. 1934). The absence of chlamydospores (htille cells ?) 
in a strain of A. sydowi is hardly a sound basis for separation. 

Sterigmatocystis cyaneus Mattlet (Ann. Soc. Belg. Med. Trop. 6: 32, 1926) was 
described without data to separate it from A. sydowi. 

S. cameleo Sartory, Sartory, and Meyer (Ann. Mycol. 29: 360-361, PI. Ill, figs. 7-8. 
1930) by description must have been some strain of A . sydowi although the very small 
smooth conidia do not agree. It may possibly represent a form near A. humicola as 
described by Chaudhuri and Sachar (see p. 193). 



THE ASPERGILLUS VERSICOLOR GROUP 187 

Aspergillus jamts Raper and Thorn, in Mycologia 36 : 55G-561, fig. 1. 1944. 

Species characterized by conidial heads of two distinct types, (1) large 
white heads borne upon long conidiophores terminating in strongly clavate 
vesicles and (2) smaller, dark green heads borne upon short conidiophores 
with typically ovate vesicles (PI. I E and F, and V B). 

Colonies varying greatty in color and in texture depending upon the sub- 
stratum and the temperature of incubation. Upon Czapek's solution agar 
at 24° C. (fig. 54 A) colonies spreading irregularly, usually consisting of a 
central floccose mass 1 to 2 mm. deep, pale yellow-buff in color, bearing few 
and scattered fruiting structures surrounded by an irregular zone of 
crowded fructifications with dark green heads occurring in a dense stand 
adjacent to the substratum (fig. 54 D) and with numerous long-stalked 
white heads projecting above this layer (fig. 54 C); reverse in dull yellow to 
light brown shades. When incubated at 20° C, colonies more restricted, 
less floccose and consisting almost exclusively of a dense stand of long- 
stalked white heads with small green heads absent or developing only in age, 
and arising from trailing aerial hyphae entwined among the white fruiting 
structures. When incubated at 30 to 32° C. colonies close-textured, pre- 
dominantly green but with central area commonly showing irregular patches 
of massed hiille cells, buff to dull yellow in color. Conidial heads abundant 
and consistently dark green in color. Reverse in dull brown shades. 

White conidial heads loose in texture (fig. 54 C), consisting of radiating 
and divergent chains of conidia, eommonfy 150 to 200m in diameter, occa- 
sionally larger. Conidiophores long, thin, mostly 2 to 2.5mm. in length by 
8 to 10.5m in diameter, occasionally larger, erect, essentially uniform in 
diameter throughout but often marked by numerous and irregularly spaced 
constrictions, walls smooth, colorless, approximately 1 to 1.4/z in thickness. 
Vesicles thin-walled, clavate (fig. 54 G), mostly 45 to 60m by 15 to 18m with 
individual structures larger or smaller, entire surface loosely covered by 
sterigmata as a rule, but often showing barren areas which may occupy any 
part of the sterigmatic surface. Sterigmata in two series, primaries 7 to 
10m by 3.5 to 4.5m; secondaries 6 to 8m by 2.5 to 3m. Conidia smooth, color- 
less, globose to subglobose, mostly 2 to 2.5m, with maximum about 2.8m. 

Green conidial heads compact, radiate when young, becoming columnar 
in age and often spreading into two divergent columns. Heads at first in 
blue to blue-green shades near dark gobelin blue (Ridgwav, PI. XXIV), 
becoming dark olive-gray in age (Ridgway, PI. LI), in size commonly rang- 
ing from 60 to 75m in diameter to 200 to 300m in length. Conidiophores 
erect, commonly 300 to 400m in length, by 6.5 to 8m in diameter, of uniform 
thickness throughout, walls smooth, colorless or very faintly green, approx- 
imately 1 to 2m thick, enlarging rather abruptly into an ovate vesicle. Ves- 
icle thin-walled, variable in form and dimensions, but commonly ovoid (fig. 




i o 



l\ 



i 



Fig. 54. Aspergillus janus; NRRL No. 1787. A and B, Colonies on Czapek's 
solution agar and malt agar, respectively, showing a, scattered white conidial heads, 
b, crowded green heads, and c, massed niille cells, two weeks old, incubation at 2-1° 

C, X i. C, Characteristic white heads borne on long thin conidiophores, X 7.5. 

D, Massed green heads in dense stand adjacent to the substratum, with white heads 
projecting from the left, and with localized development of hulle cells in two limited 
areas at right, X 7.5. E, Heads of mixed character with older (outer) conidia white 
and younger (inner) conidia green, X 24. F, Hulle cells, X 350. G, Typical white 
head showing clavate vesicle, double sterigmata and small, smooth conidia, X 450. 
H, Head of mixed character, at this stage (early) producing small white, smooth 
conidia, X 450. /, Typical green head, showing small, nearly globose vesicle, double 
sterigmata and larger, echinulate, green conidia, X 450. (Reprinted from Raper 
and Thorn, "New Species of Aspergilli from Soil," Mycologia 36: Nov -Dec. 1944.) 

188 



THE ASPERGILLUS VERSICOLOR GROUP 189 

54 I) and occasionally conspicuously elongate, typically fertile over the 
entire area, ranging in size from 20 to 30/x by 12 to 18/z. Sterigmata in 
two series, rather loosely arranged, primaries 7 to 10m by 4 to 4.5m; sec- 
ondaries 6 to 8.5^ by 2 to 2.8m- Conidia dark green in mass, conspicuously 
spinulose (fig. 54 I), globose, mostly 2.5 to 3.5/x, occasionally larger or 
smaller. 

Conidial heads of mixed character, containing both white and green 
spores, commonly encountered (fig. 54 E), usually borne upon long conidio- 
phores approaching and often equalling in length those of white heads, 
vesicles clavate (fig. 54 H), sterigmata at first bearing colorless smooth- 
walled conidia, but subsequently bearing dark green spinulose conidia. At 
temperatures of 24° C. and above, thick-walled hulle cells abundant, irregu- 
lar in form (fig. 54 F), commonly globose to subglobose, not infrequently 
elongate, commonly more or less curved and often lobed. 

Colonies upon malt extract agar growing luxuriantly (fig. 54 B), generally 
loose in texture with aerial mycelium prominent, conidial heads normally 
more abundant than upon Czapek agar, the proportion of white to green 
heads varying with the temperature of incubation. 

Colonies upon hay infusion agar spreading broadly, consisting of a thin 
submerged mycelium from which develop erect, white and green conidial 
structures, the relative proportion of these types being dependent upon the 
temperature of incubation ; since comparatively meager growth occurs upon 
this medium, and since there is a minimum of aerial vegetative hyphae, it 
constitutes a very favorable substratum upon which to observe the forma- 
tion of the contrasting fruiting structures characteristic of the species. 

The binomial Aspergillus janus was selected for this species because of the 
contrasting types of conidial heads produced — it is literally a "two-faced" 
mold. 

Type culture NRRL No. 1787 was isolated in February 1942 from 
Panama soil collected during the summer of 1941 by John T. Bonner of 
Harvard University. Three additional isolations by members of the 
Northern Regional Research Laboratory staff have since been made from 
Panama soils subsequently collected by Mr. Benjamin T. Coghill. 

It is believed that this species represents a normal component of the 
microflora of Panama. Additional evidence in support of this view is 
furnished by the fact that in 1925 Professor Roland Thaxter sent to Thorn 
under the label "white Panama Aspergillus" a representative of this species. 
The form was never described by Thaxter, and viable cultures of it were lost 
from our collection some time prior to 1930. As the correspondence of the 
time is remembered, Thaxter was plagued by the presence of a small green 
mold which repeatedly appeared in his cultures as a "contaminant." 
Fortunately, the original tube received from Thaxter has been preserved, 



190 A MANUAL OF THE ASPERGILLI 

and re-examination of this culture leaves no question but that he was deal- 
ing with a strain of the species here described, and that the green form 
which troubled him so much was not, in fact, a contaminant but a different 
phase of the same fungus. 

Aspergillus janus var. brevis Raper and Thorn, in Mycologia 36 : 561-563, 

fig. 2. 1944. 

The variety differs from the species in a number of particulars, foremost 
among which are (1) the reduced length of the conidiophores bearing both 
white and green heads and (2) a consistent tendency for white and green 
conidial structures to develop in approximately pure stands and to appear 
as contrasting radial sectors. 

White conidial heads are of the same general pattern and form as in the 
species, but are of somewhat smaller dimensions, are borne upon conidio- 
phores generally less than 2 mm. in length by 6 to 8m in diameter, and are 
characterized by elongate but not strongly clavate vesicles measuring 20 
to 25^ by 14 to 18m; conidia are smooth-walled, colorless, globose to sub- 
globose, 2.2 to 2.8m in diameter. Green conidial heads are compact, glo- 
bose to somewhat columnar, borne upon conidiophores 75 to 125m by 4 to 
6m with globose to subglobose vesicles measuring 8 to 15m by 10 to 18m; 
conidia are dark blue-green, strongly echinulate, and 3.5 to 4.5m in diameter. 

The vesicles of white heads in the variety brevis are of approximately the 
same size and form as the vesicles of green heads in the species itself, 
whereas the conidiophores bearing each type of head are approximately 
one-half the length of those bearing the same type of head in the species. 
The most striking character distinguishing the variety, however, is the 
manner in which areas of white and green heads are sharply separated along 
radial lines. Conidial heads of mixed character are produced and usually 
can be found along the frontier between white and green sectors. 

Type culture NRRL No. 1935 was isolated in July 1942 from a sample of 
soil collected in Alameda in southern Mexico, and forwarded to us in June 
by Mr. William B. Roos. 

Aspergillus versicolor (Vuill.) Tiraboschi, in Ann. Botan. (Rome) 

7:9. 1908. 

Synonym: S. versicolor Vuillemin. Mirsky, B., in These de Med. 
Nancy, no. 27, p. 15 et seq. 1903. See Thorn and Church, 
The Aspergilli, p. 142, 1926. 

Colonies upon Czapek's solution agar rather slow growing, compact, in 
some strains velvety and consisting almost entirely of closely crowded 
conidiophores arising from the substratum, in other strains showing a 
marked development of floccose hyphae bearing more or less abundant 



THE ASPERGILLUS VERSICOLOR GROUP 



191 



conidiophores as short aerial branches, in still others a combination of both 
growth types with colony centers initially floccose and outer areas almost 
velvety, at first white, passing through shades of yellow, orange-yellow, tan, 




Fig. 55. Aspergillus versicolor, strain NRRL Xo. 227. Photomicrograph showing 
details of head structure. Note particularly the double series of sterigmata, X 2100. 



to yellowish-green shades such as pea green or sage green (Ridgway, PI. 
XLVII) depending upon the strain (PI. V C and D, and fig. 53 C and D) and 
conditions of culture, occasionally with the green colors almost or com- 



192 A MANUAL OF THE ASPERGILLI 

pletely lacking; reverse and substratum occasionally colorless or nearly so, 
mostly passing through shades of yellow to orange then rose, purple-red or 
red, the particular shade and intensity of color normally persisting as a 
strain or varietal character. Heads roughly hemispherical, radiate, up to 
100 to 125m in diameter; rarely approaching columnar. Conidiophores 
colorless, smooth, up to 500 or even 700m by 5m or approaching 10/x near the 
vesicles. Vesicles 12 to 20m in diameter, fertile area hemispherical or semi- 
elliptical (fig. 55) passing almost imperceptibly into the funnel-like enlarged 
apex of the conidiophore. Sterigmata in two series, primary commonly 8 
to 10/x by 3m, occasionally less, secondary 5 to 10m by 2 to 2.5m. Conidia 
globose, usually delicately echinulate, mostly 2.5 to 3m, occasionally 3.5m or 
4m, usually borne in loosely radiating chains. 

In occasional highly colored strains vesicles and sterigmata may be 
colored. 

Hiille cells of the Aspergillus nidulans type are occasionally seen. 

Neither perithecia nor sclerotia have been found. 

The diagnosis is drawn broadly enough to cover a common and very 
abundant series of organisms which vary greatly in colony appearance. In 
some strains aerial growth is composed of conidiophores and heads only; in 
others it is made up of floccose felts or ropes of hyphae bearing short conidio- 
phores and small heads which retain the characteristic arrangement of 
vesicles, sterigmata, and conidial chains. 

The wide range of colony structure and coloration of strains, 1 and the 
varied conditions under which these have been collected probably account 
for the appearance of many names in the literature which refer to cultures of 
the group but which cannot be safely separated and identified by the de- 
scriptions given. 

The following list of species are believed to represent synonyms: 

S. ambari Beauregard (Ann. de Micrographie 10: 255-278, pi. 1. 1898). Probably 
one of the series, but never identified again. 

S. bicolor J. Ray (Rev. Gen. Bot. 9: 193-212, 245-259, 282-304, PI. 12-17. 1897) 
is probably one of this series. Primary sterigmata were reported filled with red 
coloring matter and conidia globose, spinulose, up to 2.5ju in diameter. 

Sterigmatocystis brodeni Mattlet (Name only in Ann. Soc. Beige Med. Trop. 6: 
31, 1926; discussion, ibid 4: 167-171, figs. 1, 2. 1924). Apparently a member of 
this series. 

A. flavo-viridescens Hanzawa (Jour. Coll. Agr. Tohoku Imp. Univ. Sapporo 4: 
232-3, pi. 21, figs. 1-4. 1911). Culture No. 4291.10 (Thorn) received from Hanzawa 
under this name belongs to the A. versicolor group. 

A. versicolor var. glauca Blochwitz (Ann. Mycol. 32(1/2): 86. 1934). This variety 
has the color of the A. glaucus group which is more deeply blue than A. versicolor. 
Isolated in the skin clinic at Kiel upon human skin showing "ringworm.' 



>> 



1 For a more complete discussion of the range of cultural types found in this series, 
the reader is referred to Thorn and Church, The Aspergilli, pp. 142-145, 1926. 



THE ASPERGILLUS VERSICOLOR GROUP 193 

8. glauca Bainier (Bui. Soc. Bot. France 27: 29-30, pi. 1, fig. 3, 1880, ibid. 28: 77, 
1881). From extract of henbane, dregs of wine, casks, and corks. 

A. globosus Jensen (N. Y. Cornell Agr. Exp. Sta. Bui. 315, p. 482, 1912). The 
type culture received from Whetzel (Thom No. 2705) is certainly a member of this 
series, and is characterized by yellowish-green to olive-green conidial areas, with 
colony reverse in yellowish-orange to wine red. 

S. polychroma Ferraris (Fl. Ital. Crypt. Hyph. p. 640). Syn. A. versicolor, fide 
Tiraboschi, in Ann. de Botanica (Rome) 7: 9, 1908. 

S. spuria Schroeter (Cohn Krypto. Fl. von Schlesien 3: 2 Halfte, Lief. 1, p. 218, 
1893). Position in doubt. May represent a form of A. versicolor similar to the flesh 
colored forms discussed by Thom and Church in The Aspergilli, p. 145, 1926, or may 
belong with A. carneus (See p. 201). 

A . tabacinus Nakazawa, Simo, and Watanabe (Jour. Agr. Chem. Soc. Japan 10(2): 
177-178, 1934). The detailed figures given in contrast to their own strain of A. versi- 
color showed differences which disappear when large numbers of isolations are 
studied. 

Aspergillus humicola Chaudhuri and Sachar, in Ann. Mycol. 32: 97. 1934. 

Characterization after Chaudhuri and Sachar 

Colonies on Czapek's solution agar, at first white passing through shades 
of olive-gray (Ridgway, PI. XLVI. 20. o_yy ) velvety at margin, floccose 
toward the center; reverse and substratum in shades of yellow, heads radi- 
ate. Conidiophores arising directly from the substratum, up to 300m in 
length by 4 to 5.4m in diameter, or as short branches, about 70m long, from 
aerial hyphae; walls smooth and almost colorless. Vesicles 9 to 15m in 
diameter, colorless, flask-shaped, with sterigmata radiating from the whole 
surface of the larger heads, or only borne in the upper third in small heads; 
primary sterigmata 3.6 to 5.4m by 1.8 to 2m; secondary sterigmata 3.6 by 
1.8m- Conidia globose, smooth, 2 to 3m in diameter, in radiating chains. 

Neill (1939) is believed to have correctly placed this organism with A. 
versicolor and its allies despite the smoothness of its conidia. The "almost" 
colorless conidiophore suggests relationship to Aspergillus ustus; in this 
group certain forms (e.g., Blochwitz's A. ustus var. laevis) apparently have 
spores smooth or nearly so (see p. 175). 

Pathogenesis 

Aspergillus sydowi is not reported by name as a parasite, but strains 
described in terms which must be interpreted as placing them with A. sydowi 
includes A. tunetanus (Langeron) Dodge from fleshy lesions on a hand in 
Tunis: A. Vancampenhouti (Mattlet) Dodge also from tropical Africa; A. 
cyaneus (Mattlet) Dodge from the same region. The evidence at hand 
links A. sydowi more closely with A. nidulans than was formerly supposed, 
although such relationship is strongly indicated by the identical character 
of their hulle cells. The fragmentary descriptions commonly given for 



194 A MANUAL OF THE ASPERGILLI 

individual pathogenic molds isolated by persons unfamiliar with the litera- 
ture are rarely definite enough to separate nearly related forms. 

An occasional culture of A. versicolor is obtained from apparently patho- 
genic sources. Thus far experimental work has not shown evidence of 
actual lesions in human flesh. Little colonies bearing green heads and 
conidia were drawn with a breast pump from an inflamed mammary gland 
and kept in culture for many years but failed to grow in laboratory media at 
blood heat, Aspergillus versicolor var. glauca Blochwitz was isolated from 
human skin showing "ringworm" at the skin clinic in Kiel, but apparently 
pathogenicity was not proved experimentally. Strains of A. versicolor are 
frequently observed upon dried salted lean beef, thus showing its capacity 
to grow in and upon meat products, but not giving direct evidence of par- 
ticipation in any pathological process. 

Occurrence and Economic Importance 

Members of the Aspergillus versicolor group appear widely distributed in 
soil, on spoiling and drying food stuffs, breads, cereals, old cheese, dried 
meats, cured India rubber, musty vegetable products, and other substrata 
characterized by a moderately low water content or containing factors toxic 
to most organisns. They are reported as capable of decomposing certain 
paraffins. The production of proteolytic enzymes by most strains is shown 
by the digestion of milk and the liquefaction of gelatin. Aspergillus 
sydowi, in particular, is a characteristic component of all soil examined. 

Fat production by .4. sydowi has been studied quite extensively by Pro- 
fessor Peterson and associates at the University of Wisconsin. For reference 
to this work see the various papers listed in the Topical Bibliography under 
the heading "Chemistry of Mold Tissues." 



Chapter XV 
THE ASPERGILLUS TERREUS GROUP 

Outstanding Characters 

Heads columnar, in cinnamon, pale buff or light flesh colors. 
Conidiophores smooth, colorless, rarely exceeding 250m in length. 
Vesicles hemispherical, with upper half to two-thirds covered by sterig- 

mata. 
Sterigmata in two series, generally crowded. 
Conidia smooth, globose to slightly elliptical, small. 

Included here are members of a variable and cosmopolitan group of 
Aspergilli especially common in soil. They differ markedly in color and in 
colony appearance and to a lesser degree in the texture of their conidial 
heads. The group may be separated as follows: 

Group Key 

I. Conidial heads in cinnamon or orange-brown shades, compact, uniform in di- 
ameter throughout A . lerreus series 

A. Colonies velvety, conidiophores mostly in a dense stand arising from the 

substratum. 

1 . Conidial heads in dull cinnamon shades A . lerreus Thorn 

2. Conidial heads orange-brown near xanthine orange (Ridgway) 

A. lerreus var. boedijni (Bloch.) n. var. 

B. Colonies floccose, conidial heads arising from aerial hyphae. 

1. Mycelium colorless, heads light pinkish-cinnamon in color 

A . lerreus var. floccosus Shih 

2. Mycelium yellow, heads developing late, in cream or light tan shades 

.4. terreus var. aureus n. var. 

II. Conidial heads white or flesh colored, loose textured not strictly uniform in 
diameter A . carneus series 

A. Colonies in light flesh colors, ranging from near white to vinaceous-fawn 

(Ridgway). Thick-walled hyphae suggestive of hiille cells are generally 
present A. carneus (van Tieghem) Blochwitz 

B. Colonies persistently white or becoming dull ivory in age. Thick-walled 

cells often present A. niveus Bloch. 

Aspergillus terreus Thorn, in Turesson, Gote, Svensk Botanisk Tidskrift 

10: 5, 1916, without description; diagnosis Thorn and Church, 

in Amer. Jour. Bot. 5: 85-6. 1918. 

Synonym: A. galeritus Blochwitz, in Ann. Mycol. 27(3/4): 205. Taf. 
III. 1929. 

Colonies upon Czapek's solution agar growing well at room temperature 
and up to 37° C, spreading, plane or marked by shallow radial furrows, 

195 



196 



A MANUAL OF THE ASPERGILLI 




Fig. 56. Aspergillus terreus. A, Colony margin of a typical strain, NRRL No. 
265, showing crowded, long columnar heads, X 14. B, Scattered but typical conidial 
heads of an ultra-violet light-produced mutation unable to utilize NO3 nitrogen, 
X 50. Both cultures on Czapek's solution agar, 2 weeks. 



THE ASPERGILLUS TERREUS GROUP 197 

velvety (fig. 58 A) or in some strains showing tendency toward floccosity in 
central colony areas, heavy sporing throughout with massed columnar heads 
giving to colonies their characteristic color and texture, in color ranging 
through various cinnamon shades (Ridgway, PI. XXIX) depending upon 
the abundance and maturity of the heads (PI. V E), amber exudate produced 
in some strains, odor transient to none, reverse in dull yellow to brown 
shades. Conidial heads long columnar (fig. 56 A), with conidial chains 
compacted together, of uniform diameter throughout entire length, com- 
monly ranging from 150 to 500m or more in length by 30 to 50m at maturity, 
ranging from cinnamon-buff through cinnamon to Sayal brown (Ridgway, 
PI. XXIX). Conidiophores more or less flexuous, smooth, colorless, com- 
monly ranging from 100 to 250m by 4.5 to 6.0m, approximately uniform in 
width throughout (fig. 57 A and 19 A). Vesicles hemispherical, dome-like, 
commonly 10 to 16m in diameter, merging almost imperceptibly into the 
supporting conidiophore. Sterigmata in two series, primaries crowded 
(fig. 57 A), parallel, 5.0 to 7.0m by 2.0 to 2.5m, secondaries closely packed 
5.5 to 7.5m by 1.5 to 2.0m- Conidia globose to slightly elliptical, commonly 
1.8 to 2.4m in diameter. 

Species description based upon type strain NRRL No. 255 (Thorn No. 
144) and innumerable additional isolations from soils and other sources in 
this country and abroad. The species is especially abundant in warm and 
comparatively dry arable soils. 

The great majority of isolates belonging to this series fall within A. 
terreus in its strictest sense, and duplicate in all essential particulars the 
description given above for this species. Nevertheless, wide natural varia- 
tion among strains is encountered when great numbers are isolated from 
widely separated sources. Some of these are quite striking in appearance 
and have apparently furnished the bases for species and varietal description 
by other workers. 

Aspergillus terreus var. boedijni (Bloch.) n. var. 

Blcchwitz, in Ann. Mycol. 32(1/2): 83, 1934, described Aspergillus 
boedijni as a new species differing from Aspergillus terreus primarily in the 
color of its conidia. These were reported as pure yellow at first, becoming 
pure brown or ochraceous-brown in age, and brighter than .4. galeritus 
Blochwitz. 1 In our experience strains are occasionally encountered which 
are characterized by a bright, orange-brown color instead of the dull cinna- 

1 A. galeritus Blochwitz (Ann. Mycol. 27(3/4) : 205, Taf. Ill, 1929) is a redescription 
of A. terreus Thorn. Blochwitz acknowledged having Thorn's type at hand when 
he renamed this species. No reason was given beyond the claim that he had had the 
organism in culture for some years before Thorn's description oi' A. terreus was 
published. 



198 A MANUAL OF THE ASPERGILLI 

mon shades typical of the species. Among cultures currently under exami- 
nation, this character is noted in an isolate from Argentine soil, is somewhat 
more marked in NRRL No. 680 from Dr. G. A. Ledingham, Ottawa, 
Canada, and is particularly striking in NRRL No. 1913 isolated by Dr. C 
W. Emmons from Arizona soil. In all of these strains the basic morphology 
is that of a typical A. terreus, hence Blochwitz's separation is reduced to 
varietal rank. 

Aspergillus terreus var. floccosus Shih, in Lingnan Sci. Jour. 15 : 372, PI. 16, 

fig. 3, 1936. 

Strains characterized by deep floccose colonies in which conidial heads 
are less abundant, develop late, and are borne almost entirely upon aerial 
hyphae, are frequently encountered (fig. 58 B). While the conidial struc- 
tures of certain of these strains appear entirely normal, in the majority of 
isolates the heads are somewhat less compact and generally lighter in color. 
This color difference is particularly marked among isolates from soils col- 
lected in Texas, Central America, and Cuba with color commonly light 
pinkish-cinnamon (Ridgway, PI. XXIX) to vinaceous-buff (PL XL) in age. 
No sharp line of separation can be drawn between typical strains of A. 
terreus and the floccose forms under consideration since isolates of interme- 
diate character are encountered; nevertheless, these strongly floccose 
cultures occur with sufficient frequency to warrant recognition of Shih's 
varietal designation if his interpretation is somewhat broadened. The 
variety is considered by the writers as a strongly floccose Aspergillus terreus 
in which the head is commonly less compact and lighter in color, but with 
the basic morphology of the conidial apparatus remaining that of the species 
proper. This variety is represented by such strains as NRRL Nos. 1920 
and 1921, isolated from Cuban soil contributed by Professor J. M. Osorio, 
University of Havana; No. 1922, isolated from Texas soil, collected and sent 
to us by Dr. F. E. Clark from Greenville, Texas. 

Other strains examined almost completely bridge the gap between the 
pale colored strains of A. terreus var. floccosus and the light flesh colored 
forms characteristic of Aspergillus carneus. 

Aspergillus terreus var. aureus n. var. 

This new and striking variety differs from the species in a number of par- 
ticulars: colonies upon Czapek's solution and malt extract agars are com- 
paratively slow growing, floccose, ranging up to 3 to 4 mm. deep, and are 
bright golden yellow in color. Conidial structures are produced tardily and 
in limited numbers. Conidiophores are appreciably longer than those of 
the species, often becoming 500m or more in length, and bear columnar 
heads, generally loose in texture, ranging in color from cream or light buff 



THE ASPERGILLUS TERREUS GROUP 



199 



to light pinkish-cinnamon. Microscopically, the conidial structures ap- 
proximate those of the species itself. Separation as a new variety is based 
primarily upon the characteristic coloration of the growing colony. 




Fig. 57. Conidial structures of members of the Aspergillus terreus group, X 840. 
A, A. terreus, type strain NRRL No. 255 (Thorn No. 164). B, A. terreus var. aureus, 
NRRL No. 1923. d, C 2 , and C 3 , A. carneus, NRRL No. 1928, conidial heads vary 
greatly in size. D u D 2 , and D 3 , A. niveus, NRRL No. 515, conidial heads of varying 
dimensions. 



Type strain NRRL No. 1923 (fig. 16 D) was isolated from Texas soil 
contributed by Dr. F. E. Clark. Additional strains showing approximately 
the same cultural and morphological characteristics have been isolated 
from soils collected in Arkansas and Arizona. In A. terreus var. aureus the 
yellow coloring matter is lodged in the vegetative mycelium, and there are 
no suggestions of hulle cells; in A. carneus, however, approximately the same 



200 



A MANUAL OF THE ASPERGILLI 



yellow tints are developed through the massing in localized colony areas of 
thick-walled hyphae suggestive of hiille cells (see p. 201). 

At least two other species have been described which are believed to 
represent probably synonyms of Aspergillus terreus: 

Aspergillus fuscus Anions (Archief. voor de Suikerindustrie in Nederlandsch. 
Indie Jaarg. 29, Deell, pp. 8-10, 1921) by description is obviously a form closely 
related to, if not identical with, A. terreus. 






Fig. 58. Aspergillus terreus group; different species growing upon Czapek s solu- 
tion agar at room temperature. A, Typical, heavy sporing strain of A. terreus, 
NRRL No. 265. B, A. terreus var. floccosus, characterized by loose floccose colonies 
and limited spore production. C, A. carneus, NRRL No. 1928, heavy sporing and 
characterized by flesh-colored conidia. D, A. niveus, NRRL No. 515, characterized 
by white conidial heads. 

Aspergillus cmnamominus (Weiss) Dodge, in Med. Myc, p. 627. 1935. 

Synonym: S. cinnamominus Weiss, in Ann. Parasitol. Hum. Comp. 8: 189-193, 
5 figs 1930. 
Characterization from Dodge: Hyphae septate and branched; conidiophores 
simple, 5m in diameter, vesicle 12 by 9 M . Primary phialides cylindric, 4 M long, bearing 



THE ASPERGILLUS TERREUS GROUP 201 

2 to 3, sometimes 4 secondary phialides, 5 to 6ju long. Conidia spherical, 1 to 2/x in 
diameter, brown. Other spores, borne laterally on short branches (phialides ?), 

3 to 4p. Chlamydospores occasional. Under some conditions, only monstrous 
phialides are formed, with a "pleomorphic" mycelium resulting. Described as pres- 
ent in lesions of pityriasis versicolor flava. Inoculation into the human skin repro- 
duced the original disease. From the description given the organism is some strain 
of the A. terreus group. 

Aspergillus carneus (v. Tiegh.) Blochwitz 

Synonyms: Sterigmatocystis carnea v. Tiegh., in Bull. Soc. Bot. France 
24: 103. 1877. Cited also in Saccardo Sylloge 4: 74, and 
in Wehmer's Monograph, p. 127 (Mem. Soc. Phys. Hist. 
Nat. Gen. pp. 1-157. 1899-1901). 
Sterigmatocystis spuria Schroter as a bibliographic change in 
Colin Krypt. Fl. Schleisen 3 : 2 Helfte Lief. 1, p. 218. 1893. 
Aspergillus carneus Blochwitz, in Ann. Mycol. 31(1/2): 81. 
1933. 

Colonies upon Czapek's solution agar growing well at room temperature 
spreading, plane or radially furrowed, more or less floccose, at first white 
but becoming pale vinaceous-fawn to vinaceous-fawn (Ridgway, PI. XL) 
with the development of mature fruiting structures (PI. V F, and fig. 58 C), 
ranging from 1 to 2 mm. or more deep in central areas to very thin and 
spreading at colony margin, comparatively heavy sporing throughout with 
fructifications arising from both aerial and submerged mycelium, some 
strains showing limited areas yellow in color from an underlying felt of 
heavy-walled sterile hyphae suggestive of hulle cells (fig. 49 F) ; odor often 
pronounced, somewhat putrid; reverse in orange-yellow, bright orange, to 
deep brown shades; conidial heads loosely columnar, averaging 150 to 200m 
by 25 to 35m, but commonly somewhat larger, varying in color as the colony. 
Conidiophores variable in length, mostly 250 to 400m but ranging up to 1 
mm., occasionally bearing secondary fruiting structures as short and irregu- 
larly placed branches, smooth, sinuous, uncolored, mostly 3.5 to 6.0m in 
diameter. Vesicles hemispherical, ranging from 5.5 to 9m, rarely as much as 
10m (fig. 57 Ci). Sterigmata in two series, primary 5.5 to 6m by 2 to 2.5m, 
secondary 5 to 5.5m by 1.8 to 2m, commonly very few primary sterigmata 
present. Conidia globose to subglobose, thin-walled, averaging 2.4 to 2.8m, 
with maximum rarely exceeding 3.2m. 

Colonies upon malt extract agar growing more restrictedly, heavier spor- 
ing, with pigmentation generally more pronounced and with conidial heads 
averaging slightly larger than on Czapek's solution agar, otherwise dupli- 
cating the above description. 

This species is represented by strains NRRL No. 527, isolated as an air 
contaminant in Washington; NRRL No. 298, isolated from Kansas soil; 
and other soil isolations from different parts of the United States, Mexico, 



202 A MANUAL OF THE ASPERGILLI 

Cuba, and Central America. Strains differing from the above in the 
absence of any yellow, thick-walled hulle-like cells are occasionally isolated 
from soils. NRRL No. 1928, isolated from Arkansas soil, is representative. 
In the absence of any yellow component, the color of these strains is more 
accurately described as pale to light grayish-vinaceous-fawn (Ridgway, PI. 
XXXIX). 

The name Aspergillus carneus is revived to cover the forms under con- 
sideration since their most obvious identifying characteristic is the pale 
flesh color of their massed conidial heads. It is the belief of the writers that 
van Tieghem probably had in hand some member of this series when he 
proposed the name S. carnea, although, due to the inadequacy of his descrip- 
tion, it is now impossible to establish this point with certainty. In any case, 
the name is excellently descriptive of strains commonly encountered, hence 
its application in this connection. 

In describing Aspergillus carneus as a new species, Blochwitz (1933) 
acknowledged the earlier use of this specific name by van Tieghem but dis- 
regarded its validity. He undoubtedly applied it to a member of the 
species as it is considered by us since he noted that it differed from A. 
terreus (A. galeritus) principally in the flesh to rose color of its conidia. It 
is believed that Blochwitz's A. niveus var. nubila (1934) likewise represented 
a strain of A. carneus characterized primarily by conidia of darker rose, a 
condition which in older cultures is frequently suggested by NRRL No. 
1928 cited above. 

Oilman and Abbott in their "Summary of Soil Fungi" (1927) called atten- 
tion to the repeated isolation, from Louisiana soils, of forms with the 
"general morphology of the Aspergillus candidus group but producing bright 
pink conidial heads." While the writers think the affinities of these forms 
lie more with Aspergillus terreus (columnar heads, colorless conidiophores) 
and Aspergillus fiavipes (elongate, irregular hiille cells) than A. candidus, 
we have every reason to believe they were dealing with forms similar to 
those here designated A. carneus. 

S. albo-rosea Sartory, Sartory, and Meyer (Ann. Mycol. 28: 358-359, PI. Ill, 
fig. 1-6, 1930) apparently represents a member of this series. This is indicated by 
the described coloration of colonies and more particularly by the detailed measure- 
ments cited for it. 

Aspergillus niveus Blochwitz, in Ann. Mycol. 27(3/4): 205-6, fig. 2, 

Taf. III. 1929. 

Synonym: A. eburneus Biourge, name attached to a culture received by 
Thorn (No. 5402.1 : NRRL No. 515). 

Colonies upon Czapek's solution agar white, plane or radially furrowed, 
rather slow growing, forming a dense felt of mycelium and conidiophores up 



THE ASPERGILLUS TERREUS GROUP 203 

to 600m to 1 mm. deep, thinning toward the margin (fig. 58 D) and com- 
monly spreading in unevenly radiating lines, commonly producing abundant 
amber to brown exudate ; reverse in dark yellow shades through greenish to 
brownish-black; odor slight. Conidiophores smooth with Avails colorless, 
sinuate, more or less septate, slender, 4 to 6m in diameter, enlarging to a 
hemispherical vesicle 8 to 15m in diameter or sometimes larger at the apex, 
commonly 300 to 600m in length, occasionally up to 1000m long, and on other 
substrata sometimes longer. Conidial heads showing chains of conidia in 
comparatively loose columns, most frequently 20 to 30m in diameter but in 
large heads up to 60m, with the general appearance of a snow white A. 
terreus. Vesicular area hemispherical (fig. 57 D). Sterigmata in two series, 
primary sterigmata 5 to 8m by 2.5 to 3.0m, secondary sterigmata 5 to 7m by 
2 to 2.5m- Conidia 2.0 to 2.5m, rarely more, smooth, thin-walled, colorless. 

Represented in the NRRL collection by Nos. 515 (Thorn No. 5402.1), 
288, and 1955. Repeatedly isolated from soil but less common than 
Aspergillus carneus and the ubiquitous A. terreus. 

Typically the conidial apparatus is that of a white, loosety columnar 
form of A. terreus. As described by Blochwitz, it does not show yellow in 
culture. This is true of strain XRRL No. 515, although in age this culture 
reaches dull ivory to pale buff on agar slants. In other strains, such as 
XRRL No. 1955 from Dr. Timonin in Ottawa, Canada, limited areas mav 
become yellow from the development of massed thick-walled hyphae as in 
A. carneus. 

Sterigmatocystis pusilla Peyronel (I germi atmospherici dei funghi con micelio. 
Thesis. Padova. 1931, p. 21) probably represents a synonym of A. niveus Blochwitz. 
It was described as follows: Colonies white, very thin; sterile hyphae creeping, 
sparingly branched, falsely septate, hyaline, 1.5 to 5/x in diameter; conidiophores 
erect, unseptate, hyaline, 60 to 80m by 2.5 to 3m, with apical vesicles hyaline, obovoid 
or subglobose, 7 to 10m in diameter, sterigmata radiate, in two series, with primaries 
5 to 7/x by 2 to 2.5m and secondaries 3 to 5m by 2m in groups of 2 to 3; conidia globose 
2 to 2.5m, hyaline, smooth. Habitat: from air in northern Italy at altitude of 1,700 
meters. From description this would appear to represent a short-stalked member of 
A. niveus. 

In their most typical manifestation, the conidial heads of Aspergillus 
niveus are loosely columnar; vesicles are dome-like, and fertile over the 
upper one-half to two-thirds only. They thus stand out in sharp contrast 
against the typically globose heads and completely fertile vesicles of A. 
candidus. But all heads of A. candidus are not globose, and all vesicles are 
not fertile over their entire surface. In most strains small columnar heads, 
not particularly different from those of A. niveus, can be found (fig. 60 D). 
Considering this character, together with (1) the snow-white heads, (2) the 
smooth colorless conidiophores, and (3) the small smooth conidia of both 
species, one can readily imagine that we are here dealing with two interlock- 



204 A MANUAL OF THE ASPERGILLI 

ing groups. One basic character separating the two is the production of 
sclerotia. Characteristic reddish-purple to black sclerotia are commonly 
found in white-spored strains producing wholly or in part large globose 
heads (A . candidus) ; they have never been seen in white-spored strains pro- 
ducing only loose columnar heads (A . niveus). Until additional intermediate 
forms are isolated and studied, the relationship between these white-spored 
forms must remain a matter of conjecture, although in this manual they 
are placed adjacent in what we believe to represent a natural placement of 
the different groups. 

Pathogenesis 

Strains of A. terreus grow under a wide range of temperature, including 
37° C. It is not surprising, therefore, that an occasional member of the 
series is reported as a human parasite. One of these was re-described as 
Stcrigmatocystis hortai by Langeron (1922). This culture, NRRL No. 274 
(Thorn No. 5071.1), received from France, was originally isolated from a 
human ear in Brazil. It is believed to be type and represents a characteris- 
tic strain of A. terreus. Another was found in metastatic lesions on a corn- 
husker's hand and forearm in Nebraska. Recently a strain was isolated 
from an aborted fetus from a cow in Maryland; the culture was entirely 
typical of A. terreus. 

Occurrence and Economic Importance 

Members of the Aspergillus terreus group are typically soil organisms, 
hence are most abundant in soil and upon decaying vegetation. They 
frequently occur, however, upon a great variety of materials useful to man, 
including grains in storage, straw and forage products, cotton and other 
fibrous materials not adequately protected from excessive moisture, etc. 
Aspergillus terreus and Aspergillus carneus are especially widespread in warm 
arable soils, and have been isolated in great abundance from soils collected 
in southern and southwestern United States. They are generally less com- 
mon in forest than in cultivated soils, and are rarely found in acid forest 
soils from the colder temperate zone. There is little evidence that these 
forms are especially active agents of decay, but their great abundance in 
nature indicates that they undoubtedly play a significant role in the slow 
decomposition of organic materials. Aspergillus terreus and A. carneus 
grow well at temperatures of 35 to 37° C, a character which possibly 
accounts for their great abundance in southern soils and their relative scar- 
city in soils from northern areas. 

Aspergillus terreus has become of special biochemical interest since the 
discovery in 1939 by Calam, Oxford, and Raistrick that certain strains of 
this species are capable of producing itaconic acid from sugars. Extensive 



Plate VI 

A (upper left), Aspergillus Candidas Link, XRRL No. 308. B (upper right), Aspergillus niger group, strain 
NRRL No. 67. C (center left), Tan-spored mutant of strain 67 produced bv ultraviolet radiation. D (center 
" g D d' As P er illus arenaceous Smith, NRRL No. 517. E flower left ), Aspergillus alhaceus Thorn and Church. 
XRRLN'o. 315. F (lower right), Aspergillus wentii Wehmer, NRRL No. 375. All cultures growing upon 
Czapeks solution agar. (Color photographs by Haines, Northern Regional Research Laboratory. Repro- 
duced through co-operation of Chas. Pfizer & Co., Inc.) 



THE ASPERGILLUS TERREUS GROUP 205 

investigations on this fermentation have been conducted in the Fermenta- 
tion Division of the Northern Regional Research Laboratory, and papers 
reporting these studies are currently in press. More than 300 strains have 
been tested and the most productive cultures selected for intensive study. 
Improved nutrient solutions have been developed and critical environmen- 
tal factors explored with the result that substantial yields of itaconic acid 
are now obtained. It is believed that this process may become of industrial 
importance within a reasonable period of time. (See Lockwood, Raper, 
Moyer, and Coghill; Lockwood and Reeves; Lockwood and Ward; Moyer 
and Coghill; and Raper, Coghill, and Hollaender, in the Topical Bibliog- 
raphy under "Itaconic Acid.") 

Timonin (1942) reported the production of citrinin by a white-spored 
Aspergillus identified by him as one of the A. candidus group. Careful 
examination and comparison of his culture with representative strains of 
A. candidus, A. carneus, and A. niveus show its true relationship to be with 
A. niveus in the terreus group, as it is considered here, rather than with A. 
candidus. 



Chapter XVI 
THE ASPERGILLUS CANDIDUS GROUP 

Outstanding Characters 

Conidial heads persistently white or becoming yellowish cream in age; 

typically globose, but approaching columnar in small heads. 
Conidiophores smooth, colorless or slightly yellowed in terminal areas. 
Sterigmata in two series, with primaries often much enlarged, sometimes 

varying greatly in size within the same head. 
Conidia globose or subglobose, smooth. 
Sclerotia present in some strains, dark, approaching purple to black 

when mature. 

Grouping by color lead Thorn and Church (1926) to establish their Section 
IX, or the so-called "White-spored Aspergilli." Only vaguely did they 
indicate that they had included a heterogeneous lot rather than described 
a natural group. Further study of the strains included in the "white" 
section showed the tardy development of colors approaching avellaneous 
or even carneus. The continued comparison of great numbers of strains 
in all groups has revealed such wide variations in color that the authors 
have come to regard whiteness, or lack of color, as a character of secondary 
importance in the allocation of strains to particular groups. This view- 
point is supported by the appearance, under controlled conditions, of 
white variants or mutants in a number of colored series. Yuill (1939) 
observed and isolated such colorless mutants from Aspergillus fumigatus 
and A. nidulans; Steinberg and Thom (1940) reported the same type of 
mutant for the former species; while Raper, Coghill, and Hollaender (in 
press) have succeeded in producing white mutants in Aspergillus terreus 
by irradiating spores with ultra-violet. Colorless members of the Asper- 
gillus glaucus group, represented by A. niveo-glaucus, have been isolated by 
Blochwitz, Thom and Raper, and other investigators. Long before the 
work of Yuill, Schiemann (1916) had developed two color variants of 
Aspergillus niger, A. schiemanni (Schiemann) Thom (1926, p. 172) 
and A. cinnamomeus Schiemann (1912) which differed only from the 
parent strain by a progressive reduction of the amount of coloring 
substance, presumably the aspergilline of Linossier (1891). Steinberg 
and Thom (1940), working with a strain of A. niger, again produced 
variants approximating those of Schiemann, and Whelden (1940) secured 
the same by irradiating spores of A. niger with cathode rays. In cultures 

206 



THE ASPERGILLUS CANDIDUS GROUP 207 

collected from nature from world-wide sources, strains characterized by 
heads approaching white are occasionally observed in other groups. It is 
apparent, then, that the capacity to produce a coloring substance, while 
ordinarily inherited or passed on in successive colonies of an organism, is 
not always uniformly maintained. 

In the present treatment, the writers have sought to include within the 
Aspergillus Candidas group only such forms as are clearly and closely related 
to it. The group is thus limited, essentially, to a single series containing 
only one clearly definable species, A. candidus. Different isolations vary 
materially in their general cultural appearance and in the details of their 
microscopic structure. Nevertheless, all possess the typically globose, 
white to dull buff or light gray conidial heads, the smooth colorless coni- 
diophores, and the small, smooth, colorless conidia. 

To facilitate recognition of members of the Aspergillus candidus group 
and to assist in the proper assignment of other white or light-colored 
species and strains, a general key covering all of these forms is presented. 

GENERAL KEY OF WHITE ASPERGILLI 

A. Heads (large ones) globose or radiate; conidiophores smooth-walled, colorless or 

yellowed toward the vesicle only; sclerotia occasionally seen A. candidus group 

B. Heads white, hemispherical to columnar; conidiophores smooth-walled, colorless 

A. niveus series, see p. 202 

C. Heads initially white, tending to be columnar; conidiophores smooth-walled, 

showing some shade of yellow; contorted hulle cells usually found 

A. flavipes, series, see p. 179 

D. Heads white, borne upon long, smooth-walled, colorless conidiophores terminat- 

ing in clavate vesicles, sterigmata in two series 

White-spored phase of A. janus, see p. 187 

E. Strains of white Aspergilli possessing the basic characters of their colored counter- 

parts also occur as mutations in the A. fumigatus, A. nidulans, A. terreus, and 
glaucus groups. 

Aspergillus candidus Link, Obs. p. 16, 1809. Thorn and Church, The 

Aspergilli, p. 157. 1926. 

Colonies upon Czapek's solution agar persistently white, or becoming 
cream or yellowish-cream in age (PI. VI A and fig. 59), often thin, veg- 
etative mycelium often largely submerged, surface growth usually con- 
sisting of conidiophores and heads, and with scanty sterile mycelium or 
anastomosing ropes of hyphae bearing short-stalked fruiting structures; 
sclerotia produced in occasional strains; reverse usually uncolored. Heads 
white, globose, radiate, varying in the same culture from large globose 
masses 200 to 300m in diameter to small heads often less than 100/x in 
diameter, commonly more or less elongated in heads with incomplete de- 
velopment of sterigmatic surface. Conidiophores varying with the strain, 



208 



A MANUAL OF THE ASPERGILLI 



in short or dwarf races less than 500m long, in other strains ranging up to 
500 to 1000m or longer, varying from 5m in diameter in dwarf forms to 10 
to 20m in long-stalked forms, with walls thick, smooth, colorless or slightly 
yellowed near the vesicle in certain strains in age. Vesicles typically 




Fig. 59. Aspergillus candidus. A, Strain XRRL No. 305 on Czapek's solution 
agar at room temperature, 10 days. B, Strain NRRL No. 314 on Czapek's solution 
agar at room temperature, three weeks. Note contrast between compact colony 
of No. 305, consisting of crowded conidiophores, and loose spreading colony of No. 
314 in which production of conidial heads is very irregular. C, Strain NRRL No. 
312, portion of colony showing scattered black sclerotia, X 3. D, Strain NRRL 
No. 308, conidial heads, X 18. 



globose, ranging from 40m in diameter in very large heads (fig. 60 A), and 
typically fertile over the whole surface, to small globose heads (fig. 60 B), 
often very much reduced to support simple groups of sterigmata appearing 
almost penicillate (fig. 60 D). Sterigmata typically in two series, usually 
colorless, primary varying greatly in different strains, in different heads of 




Fig. 60. Conidial structure of Aspergillus candidus, X 900: A, Large globose head 
showing large primary sterigmata, strain NRRL No. 312; B, Smaller globose head 
showing small primary sterigmata, strain NRRL No. 308; C, Head showing primary 
sterigmata of variable size, some septate, strain NRRL No. 312; D, Diminutive 
head, same strain. Great variation in dimension is characteristic of the fruiting 
apparatus of most A. candidus strains. 

209 



210 A MANUAL OF THE ASPERGILLI 

the same strain, and occasionally in the same head (fig. 60 C), ranging from 
5/i in length in some cases to 15 or 20/t and even 30/x under other conditions, 
commonly septate; secondary sterigmata usually uniform in all heads, 
from 5 to 8/x by 2 to 2.5 or 3/x. Conidia colorless, globose or subglobose 
in most strains to elliptical or barrel-form in others, thin-walled, 2.5 to 
3.5ju or occasionally 4/x, smooth. 

Reddish-purple to black sclerotia, consisting of thick-walled, parenchyma- 
like cells, occur in many strains (fig. 59 C). 

As there is no possibility of determining which of the white Aspergilli 
was in Link's possession, the name is here used to cover a whole series of 
strains which are found everywhere but are most frequent in the later 
stages of decay in vegetation and are especially characteristic of moldy 
grain. Included within the series as we consider it are two rather different 
cultural entities. The first of these is characterized by thin colonies in 
which the mycelium is largely submerged and only fruiting structures, 
commonly arranged in concentric zones, rise above the level of the sub- 
stratum (fig. 59 B). In other strains, colonies are rather floccose and some- 
what felted and often attain a depth of 2 mm., with fruiting structures 
arising from aerial as Well as submerged mycelia. Sclerotia, generally in 
purple or black shades, regularly and consistently develop in many strains 
including representatives of both of the above colony types. Conidiophores 
vary greatly in size and characteristically reach their greatest dimensions 
upon the drier portion of agar slants, or upon slightly moistened grain and 
other comparable products low in water content. The sterigmata in 
organisms of this group warrant particular attention since the two series 
commonly, but not consistently, differ tremendously in size. In many 
strains the primary sterigmata are characteristically wedge-shaped and 
reach dimensions of 25 to 30m by 10 to 12/* (fig. 60 A and C) ; such structures 
are commonly septate. In other heads from the same culture the primary 
sterigmata may be relatively small and measure 6 to 8/x by 2.5 to 3. 5/i 
(fig. 60 B). Secondary sterigmata are consistently small and of the dimen- 
sions indicated in the species description. 

"White" Aspergilli regularly constitute a normal element in the micro- 
population from moist or improperly dried grains and of comparatively 
dry vegetation undergoing slow decay. From such material, many in- 
vestigators have described molds characterized by white heads which 
obviously belong in this group, but without supplying sufficient critical 
data to permit subsequent verification of the exact types under study. 
Some of these descriptions were based upon molds growing in culture; 
more of them were not. A few of the more tangible of these probable 
synonyms will be briefly considered in this connection ; others will be found in 
the general species index (pp. 331-?). 



THE ASPERGILLUS CANDIDUS GROUP 211 

S. alba Bainier (Bull. Soc. Bot. France 27: 30. 1880) was isolated from oatmeal, 
and while incompletely described, obviously represents a member of the A. candidus 
series. Several publications present elaborate comparative tables to separate strains 
accepted as A. Candidas and A. alb us, but the many strains obtainable vary into each 
other so completely that little or no basis for separation exists in fact. 

A. albus Wilhelm (Beitr. z. Kenntn. d. Pilzgattung Aspergillus, Inag. Diss. Strass- 
burg, p. 69, 1877) was described with characters which clearly ally it with A . candidus 
but without sufficient differences to separate it from other members of this group. 

S. blanc-jaune Bainier nomen nudum — A culture from Bainier's collection received 
by Thom under this name (No. 4640.490) represents a somewhat diminutive but other- 
wise typical member of this series. 

S. albo-lutea Sartory and Meyer (Cited by Blochwitz in Ann. Mycol. 31: 73. 1933). 
Conidia were reported as turning yellowish in age. This character is common to 
many members of the group and has been so noted by Wehmer (1889-1901), Thom and 
Church (1926), and others. Retention of the species name is not warranted. 

A. basidiosepla Sartory, Sartory and Meyer (Ann. Mycol. 27: 317-320, PI. 7. 1929) 
apparently represents a member of the A. candidus series with comparatively long 
(28 to 30m) primary sterigmata which in age are characteristically septate. This char- 
acter, which appears also in some members of the A. niger and A. ochraceus groups, 
however, is not sufficiently unique to warrant specific separation. 

A. niveus var. major Blochwitz (Ann. Mycol. 32(1/2): 86. 1934). Described as 
showing vesicles globose, rarely oboval, or pear-shaped entirely covered with radiat- 
ing sterigmata which are rarely absent toward the base; closely growing conidio- 
phores 2 to 2.5 mm. high. These characters suggest relationship with A. candidus 
rather than A. niveus. 

A. okazakii Okazaki, in Centralb. f. Bakt. etc., 2 abt., 19, p. 481-484, taf. I. 1907; 
see also Centralb. f. Bakt. etc., 2 abt., 42, p. 225. 1914. This is cited by Saccardo 
in Syll. 22: 1260. 1913, as S. okazakii Saito but apparently without adequate ground 
for attributing the name or description to Saito. 

Colonies described as white to sulphur yellow; conidiophores hyaline, straight or 
sinuate, smooth or asperulate, 200 to 500m by 8 to 12m figured as undulate, especially 
toward the base, with walls 2 to 3m thick; heads 80 to 100m in diameter; vesicles 12 to 
40m in diameter; primary sterigmata 15 to 20m by 6 to 8m, secondary 8 to 14m by 2.5 to 
4m; conidia globose, hyaline, 2.5 to 5.4m, smooth, with connectives. In the event that 
continued study of these white forms reveals the existence in nature of strains with 
more or less roughened conidiophores and conidial heads ranging to yellows, recogni- 
tion of A. okazakii as a separate species would be warranted. Based upon current 
information, however, we believe it preferable to consider it synonymous with A. 
candidus Link. 

A. sachari of Chaudhuri and Sachar (Ann. Mycol. 32: 95. 1934) is more or less 
arbitrarily left where the authors put it — as one of the A. sulphureus series near A. 
quercinus in the A. ochraceus group. The heads are pale yellow, the sclerotia are 
near the colors of that group, but the conidiophore is described as colorless and 
smooth which would put it in A. candidus. 

A. sterigmatophorus Saccardo, in Mycologicae Venetae Specimen. Atti d. Soc. 
Ven. Trent, d. Sci. Nat. 2, fasc. 2: 232. Tab. XVII, fig. 5-8. 1873; Syn. S. italica 
Sacc. in F. italici no. 109, 1881 ; changed to S. italica Sacc. as a note only in Michelia 
1:91. 1877; Latin diagnosis of S. italica in Saccardo Sylloge 4: 72. 1886. Described 
from decaying corn kernels (Zea mays): white, sparse, with conidiophores un- 
branched, 2 to 3 septate above; with vesicles globose; sterigmata described as dicho- 
tomously or trichotomously branched with ultimate cells bearing conidial chains; 



212 A MANUAL OF THE ASPERGILLI 

conidia globose about 6^ in diameter, with connectives. The description repeats 
observations as to occurrence and appearance that are frequently seen. No one 
has since reported a member of the A. candidus group with conidia 6m in diameter. 
Whether the organism described by Saccardo was a large-spored mutant, not since 
isolated, remains open to question. 

OTHER WHITE ASPERGILLI 

Other Aspergilli characterized by white heads but differing basically 
in morphology from the A. candidus group occur in the Aspergillus glaucus, 
A. nidulans, A. fumigatus, and A. terreus groups. Except for an absence 
of spore color, these duplicate the morphology of the groups to which 
assigned. In fact, in all cases except that of A. niveo-glaucus in the A. 
glaucus group, they represent colorless mutations produced experimentally 
from typical parent strains (Yuill, 1939; Steinberg and Thorn, 1940; Raper, 
Coghill, and Hollaender, in press). While A. niveo-glaucus was isolated 
from nature and hence its parentage is not known, it is suspected that 
this represents a mutation of some form close to Aspergillus echinulatus. 
A. halophilus of Sartory et al (Ann. Mycol. 28: (3/4) pp. 362-3, PI. 3, 1930) 
similarly belongs in the A. glaucus group. Attention has been called earlier 
to the fact that A. candidus differs from A. niger primarily in the absence 
of color and in possessing smooth spores. The question may arise whether 
we are not here dealing with a whole series of mutations from colored forms. 
While this is possible, no proof is at hand. The fact that they constitute 
such a typical and abundant element of the micropopulation of soil, decaying 
vegetation, etc., demands that they be considered along with other major 
groups of the Aspergilli quite aside from any questions of possible origin. 

Sclerotia 

In this arrangement of the Aspergilli, sclerotia, as compact globose or 
subglobose bodies composed of thick-walled pseudo-parenchyma, are not 
found in the groups characterized by the production of perithecia and 
ascospores, and only rarely, if at all, in groups characterized by the presence 
of hulle cells. In the great groups beginning with A. candidus, sclerotia 
appear with sufficient frequency to be morphologically significant as 
indicative of class relationship. Fundamentally, the typical A. candidus 
strain differs little from the black Aspergilli except for the absence of the 
dark color and rough spores. 

Group Relationships 

While there is much evidence of relationship with the black Aspergilli 
(smooth-walled conidiophores, globose vesicles and heads, and the presence 
of sclerotia), there are also certain indications of relationship to Asper- 
gillus niveus. Typically both are characterized by snow-white conidial 



THE ASPERGILLUS CANDIDUS GROUP 213 

heads, and in all strains of Aspergillus candidus there are more or less abun- 
dant small heads which bear few and loosely arranged sterigmata in a 
manner strongly suggestive of typical conidial structures of A. niveus. 
Although it is our belief that these similarities in structure do not of 
necessity reflect close relationship between A. candidus and A. niveus, we 
do feel that there is need for additional study of strains which appear to be 
more or less transitional between the two groups. 

Occurrence and Economic Importance 

Members of the Aspergillus candidus group are very widely distributed 
in nature and occur with reasonable frequency upon vegtation in the later 
stages of decay. They are especially common upon moldy grains and 
are obviously able to grow in the presence of a very limited amount of 
moisture. 

The biochemical and physiological activities of these form have not been 
studied extensively. A. okazakii was employed by Okazaki (1907 and 
1914) for the production of a proteolytic enzyme preparation, "digestin," 
and is the basis of a Japanese patent, No. 11461, covering this process. 
Recently Timonin (1942) has employed a strain reported as belonging to 
the A. candidus group for the production of citrinin. Upon examination, 
however, this strain is found more nearly to represent A. niveus than A. 
candidus in the sense it is considered here. 



Chapter XVII 
ASPERGILLUS NIGER GROUP 

Outstanding Characters 

Conidial bonds carbon black, brownish-black or purple-brown; in mutants, 

shading toward colorless but not actually white. 
Heads typically large and globose; but small heads produced in some 

strains, in extreme cases consisting of only a few sterigmata and chains 

of eonidia. 
Conidiophores smooth, colorless or tinged with yellow --brown colors in 

the upper one-third or less, splitting lengthwise into strips and shreds 

when broken. 
Vesicles globose in large heads, fertile over the entire surface; in small 

heads often reduced to dome-like apices of short conidophores ("fumi- 

gatifonn")- 
Conidia rough, mostly showing bars or bands of brown-black coloring 

matter. 
Sclerotia characteristic of many strains, more or less irregular, ranging 

from buff through gray to almost black. 

The designation "Aspergillus niger" 1 is commonly used to cover a great 
aggregate of Aspergilli differing in details of morphology, but having in 
common the production of conidial heads which are black, brownish-black, 
purplish-brown, or in some strains lighter in color but retaining the general 
appearance oi the group. 

When examined by the hundred as they are isolated from natural sources, 
the vast majority of cultures studied show the general morphology of 
Aspergillus niger van Tieghem. if a reasonable allowance be made for strain 
variation which is characteristic of all of these groups of cosmopolitan molds. 
At the same time, other organisms scarcely distinguishable in general 
cultural appearance often show marked differences in microscopic char- 
acters, hence have formed the basis of species descriptions. Other species 
have been segregated by various authors in the belief that they bore an 
obligate relation to the particular substrata from which isolated (e.g.. 
.4. strychni. A. ficuum), but such specificity has not proved dependable. 
There are. in addition, a considerable number of probable variants whose 

1 Biourge in his last manuscript proposed the use of the name Pulli for the black 
Aspergilli in recognition of the belief that Mieheli J~29^ had one of them before him 
as his "Aspergillus capitolus capitulo pullo." 

214 



ASPERGILLUS NIGER GROUP 215 

origin is not known, and which may or may not have genetic connection, 
but which do show superficial resemblances a1 least. Some of these have 
been described as species and can be more or less readily identified. Al- 
together, there is a considerable Lisl of names which, at one time or another, 
have been applied to the black Aspergilli. It is now quite impossible to 
interpret many of these descriptions, or to find out exactly what type of 
organisms the describers had under observation. Realizing the futility 
of attempting to separate all of these species, we have endeavored to in- 
clude in the group key only such forms as possess well-marked character-, 
and those which the mycologist may reasonably expect to encounter in 
laboratory culture. 

Group Key 

I. Sterigmata in two scries. 

A. Conidia mostly less than 5m in diameter Aspergillus niger series 

1. Conidia strongly colored, rough, definitely marked with echinulae, 

tubercles or color bars. 

a. Primary sterigmata mostly under 20m in length. 

1/ Colonies with a penetrating actinomyces-like odor 

A.foetidus n. sp. <= A. aureus Xak.) 
2.' Colonies without such odor. 

a. Colonies black or deep brown to black. 

1." Heads abundant over the entire colony 

A. awamori Xakazawa 
2." Heads few, scattered at margins of colonies 

A. miyakrensis Xakazawa 
b.' Colonies yellow-brown 

Occasional Isolates. See also A. wentii group. 

b. Primary sterigmata mostly 20 to 30m long 

A. niger van Tieghem series and species. 

c. Primary sterigmata mostly 40 to 60m long 

A. phcenicis (Cda.j Thorn 

d. Primary sterigmata up to 100 to 120m long 

A. pulverulentus (McAlpinej Thom 

2. Conidia with color almost suppressed or diffused, leaving smooth walls 

with colorless spinules. 

a. Conidia almost colorless, in mass very pale cimmamon 

A. niger mut. cinnamomeus n. comb. 

b. Conidia with color diffused, sometimes a trace of roughening 

A. niger mut. Schiemanni n. comb. 

B. Conidia more than 5m in diameter Aspergillus carbonarius series 

1. Primary sterigmata less than 20m in length. 

a. Conidia purplish-black, 6 to 10m in diameter 

A . atropurpureus Zimmerman 

b. Conidia brown, 5 to 7 or 8m in diameter. .A. fumaricus Wehmer 

2. Primary sterigmata 20 to 45m in length. Conidia 6 to 8m in diameter 

A.fonsecaeus n. sp. (= S.fusca Bainierj 

3. Primary sterigmata up to 100 or 120m. Conidia 5.5 to 10.5m 

A. carbonari u.s_ fBainier) Thom 



216 A MANUAL OF THE ASPERGILLI 

II. Sterigmata in one series (Secondary sterigmata occasional in some strains) 

Aspergillus luchuensis series 

A. Colonies black or black-brown. 

1. Sterigmata 6 by 3/* (very short) A. luchuensis Inui 

2. Sterigmata about 15 to 20/xconidia 3 to 3.5yu.. .A. nanus Montagne 

and/or A. subfuscus Johan-Olsen 

B. Colonies in reddish-brown shades. 

1. Conidia globose A. japonicus Saito 

2. Conidia elliptical A. violaceo-fuscus Gasperini 

Because of their great abundance in nature, the series most closely- 
related to and including van Tieghem's species, based upon strains which 
satisfy his original description in a somewhat broadened sense, will be 
discussed first. 

ASPERGILLUS NIGER SERIES 

Species Characterized by Comparatively Small Primary Sterigmata and 

Small Conidia 

Aspergillus niger van Tieghem, in Ann. Sci. Nat. Bot., s. 5, t. 8, p. 240. 

1867. 

Synonym: Sterigmatocystsis nigra van Tieghem, in Bui. Soc. Bot. 
France 24: 102-103. 1877. See also Thorn and Currie, Jour. Agr. 
Res. 7: 1-15. 1916; and Thorn and Church, The Aspergilli, p. 167. 
1926. 

Characterization: Colonies rapidly growing with abundant submerged 
mycelium, colorless, or in some strains with more or less yellow color in 
the hyphae and in the substratum, with aerial hyphae usually scantily 
produced, but abundant in age in certain strains. Conidial heads fuscous, 
blackish-brown, purple-brown, in every shade to carbonaceous black 
(PI. VI B), varying in intensity with the quantity of coloring matter pro- 
duced; typically globose or radiate (fig. 63 C), commonly up to 300, 500, 
or occasionally 1000m in diameter with periphery variously splitting into 
radiating columns of conidia; small heads, more or less columnar and 
consisting of a few conidial chains often borne on trailing hyphae or short 
conidiophores near the substratum. Conidiophores mostly rising directly 
from the substratum, uncolored or yellow to brown near the vesicle only, 
smooth, with walls thick, frequently uneven on the inner surface and split- 
ting lengthwise into strips when broken (fig. 64 B and C), unseptate or 
with occasional thin septa, varying greatly in length and diameter in 
different strains and in colonies on different media or even in sections of 
the same colony, thus ranging from strains with conidiophores 200 to 400m 
by 7 to 10m to forms with conidiophores several millimeters long and 20m 
or more in diameter. Vesicles globose or subglobose, thick- walled, com- 









Fig. 61. Aspergillus niger group: cultures growing upon Czapek's solution agar 
at room temperature, 10 days. A, A. niger, NRRL No. 334, typical strain. B, A. 
niger mut. schiemanni characterized by colonies light brown in color. C, A.foetidus, 
NRRL No. 341, characterized by a yellowish vegetative mycelium and a strong 
actinomyces-like odor. D, A . niger, NRRL No. 346, characterized by the production 
of abundant sclerotia. E, A. phoenicis, NRRL No. 1956, characterized by long 
uncrowded conidiophores. F, A. riolaceo-fuscus, NRRL No. 360, characterized by 
compact, close-textured colonies and small heads with uniseriate sterigmata. 



217 




Fig. 62. Conidial structures in the Aspergillus niger group, X 500: A, A. niger, 
NRRL No. 326, typical head showing globose vesicle and primary sterigmata about 
twice the length of secondaries; B, A. phoenicis, NRRL No. 1956, characterized by 
long and occasionally septate primary sterigmata; C, A. carbonarius, NRRL No. 
369, characterized by la^rge primary sterigmata and large conidia; D, A. violaceo- 
fuscus, NRRL No. 360, characterized by small heads and sterigmata in a single series. 

218 



ASPERGILLUS NIGER GROUP 219 

monly 20 to 50m, occasionally up to 100m in diameter (fig. 62 A), colorless 
or more commonly more or less intensely yellow-brown. Sterigmata in 
one series in young colonies and in small heads, but typically in two series, 
colorless at times, usually more or less intensely brown, even carbonaceous, 
primary sterigmata closely packed, covering the vesicle, varying greatly 
in size in the same colony but usually 20 to 30m in length by 6 to 8m in 
diameter at the outer end; secondary sterigmata more uniform, ranging 
usually from 6 to 10m by 2 to 3m (fig. 62 A), both series often more or less 
brown to almost black. Conidia globose when ripe, with walls at first 
smooth with diffused brown or fuscous color (Ridgway, PL XLVI), then 
rough or spinulose from coloring substance deposited as tubercles, bars 
or loops between the outer primary wall and the inner, or secondary, wall, 
mostly 2.5 to 4m, occasionally up to 5m in diameter. 

Sclerotia globose, superficial, regularly produced by certain strains 
(fig. 61 D), sporadically by some, and not found in many others. 

A. niger approximating the description of van Tieghem furnishes the 
most common morphological entity among the black Aspergilli. A few 
of the substrata and locations found in our record include chronic irritants 
in the human ear, pin-point colonies in the human lung, spoiling raw sugar, 
rancid butter and other fats, floating and submerged mycelium in many 
chemical solutions. It is abundant in soil cultures from every part of 
the world, and apparently especially so in the tropics. Molds under this 
name have been used in literally hundreds of biochemical investigations. 

The introduction of a complete description from culture for each member 
of the series typically represented by van Tieghem's A. niger, but which 
vary from it in detailed measurements, calls for repetition of many common 
characters. In place of such descriptions the names and citations of the 
forms selected, either as unique or as representing sections of the series 
often encountered, are presented with the more important differences 
which furnish the bases of separation. 

Aspergillus foetidus n. sp. 

Synonym:^, aureus Nakazawa, in Inst. Gov't. Res. Formosa, Rept. 

Vol. 1, 1907. 
Not:i4. aureus Berkeley, in English Flora Vol. 5, p. 346. 1836. 
Not S. aurea Greco, in Origine des Tumeurs et Mycoses 

Argentines, Buenos Aires, pp. 671-694, fig. 418-428. 1916. 

Colonies upon Czapek's solution agar rather slow growing, producing a 
floccose basal mat of mycelium with abundant but uncrowded black heads 
above a mass of pale orange mycelium which is deeply orange in reverse. 
Heads up to 225m in diameter are borne on conidiophores about 500m 



220 A MANUAL OF THE ASPERGILLI 

long; vesicles comonly 20 to 30m in diameter, occasionally much larger. 
Primary and secondary sterigmata are both 7 to 10m by 2 to 4m; conidia 
are globose, spinulose, up to 4 or 4.5m- 

Colonies have a penetrating actinomyces-like odor (also like Penicillium 
biforme noted in Thorn, The Penicillia, p. 320. 1929), unlike any other 
species of Aspergillus. Numerous experiments over several years failed 
to justify the belief that the culture was contaminated with some actino- 
mycete. The species is known only from Nakazawa's isolates which have 
maintained the odor and orange color for many years. 

In the Awamori fermentation, A. foetidus (A. aureus Nakazawa) was 
j8, the unfavorable organism which gave a yellow color to the "Koji" used. 
Nakazawa, Simo, and Watanabe (Jour. Agr. Chem. Soc, Japan, No. 144, 
pp. 931-974, illustr., 1936) listed five varieties of A. aureus as follows: 
var. minor, var. murinus, var. acidus, var. pallidus, and var. brevis. 

Aspergillus awamori Nakazawa, in Inst, of Gov't. Res. Formosa, Rept. 

Vol. 1, 1907 and Vol. 2, 1912. 

The measurements given are only slightly different from those of A. 
foetidus ( = A. aureus). The unique odor and the yellow color in the 
mycelium and substratum were lacking. Colonies blackish-brown (deep 
chocolate) when heads were fully developed, conidiophores 1 to 2.5 mm. 
by 9 to 15m; vesicles globose, 30 to 45m hi diameter; primary sterigmata 
9 to 12m by 3.5 to 5.5m, and secondary 4.5 to 8m by 1.5 to 3.5m; conidia 
globose or somewhat elliptical 3 to 5m in long axis, fairly spinulose. 

Nakazawa, Simo, and Watanabe (Jour. Agr. Chem. Soc. Japan, No. 144, 
pp. 931-974, illust., 1936) studying the fermentation industries of Formosa 
found two general types of Aspergilli in the Awamori fermentation, a and 
(3. Type a was A . awamori for which they described the following varieties : 
var. minimus, var. piceus, var. ferrugineus, var. fuscus, var. fumeus. This 
"awamori" series produced the more desirable type of product; citric acid 
was present, as well as alcohol due to the yeast used in the inoculum. 

Aspergillus niger var. fermentari us Nakazawa, Simo and Watanabe, in Jour. Agr. 
Chem. Soc. Japan, 10(2) 1934. pp. 171-172, summarized on p. 184. Reported as a 
variety with "conidiophores 1037 to 2438m by 13.1 to 16.0m; vesicles 22.6 to 73.6m; pri- 
mary sterigmata 12.7 to 16.1m by 3.3 to 7.2m, secondary 6.8 to 9.8m by 3.3 to 4.6m; and 
conidia globose 2.3 to 4.6m in diameter." This form obviously belongs close to A. 
awamori. 

Aspergillus miyakoensis Nakazawa, Simo, and Watanabe, in Agr. Chem. 
Soc. Japan, Jour. 12(9): 963-4, fig. on 973. 1936. 

The colonies figured by the authors show a cottony mycelium with 
long-stalked heads in a broad zone near the margin. The measurements 



ASPERGILLUS NIGER GROUP 



221 



differ from ^4. foetidus as follows: primary sterigmata are reported to be 
12 to 20/x by 4.4 to 9m in contrast to secondaries 7 to 10/x by 2.5 to 5/x, whereas 




Fig. 63. Conidia, Aspergillus niger group. A , Single long chain of conidia, X 1000 
(Photograph by Edward Yuill). B, Conidia of a typical strain of A. niger, NRRL 
No. 344, X 700. C, Conidia of the citric and gluconic acid producing strain, NRRL 
No. 67, X 700. The large size and coarsely roughened walls are characteristic of 
the conidia of the latter strain. 

in A. foetidus both series measure 7 to 10/x by 2 to 4m; conidia are globose 
3.7 to 5.6m in diameter. 

The species repeats the colony appearance of certain mutants produced 
by means of chemical stimulants by Thorn and Steinberg (1939) from the 
latter 's standard strain of A. niger. 



222 



A MANUAL OF THE ASPERGILLI 



.4. hennebergi Blochwitz, in Ann. Mycol. 33: 238-9. 1935. 

Colonies described as showing the colors and general aspect of ,4. tamarii or A. 
wentii but with conidiophores browned as in the upper part of the conidiophores of 
the A. niger group, and with "red" sclerotia; relationship doubtful. See also the 
A. wentii group. 

Aspergillus niger van Tieghem, in Ann. Sci. Nat. Bot., s. 5, t. 8, p. 240. 1867. 

Van Tieghem 's strain is not fully verifiable among organisms now main- 
tained in culture, although Biourge (personal communication) believed he 
had it. We believe the name can be most appropriately used to cover 



1 




4, < v •*> t 






A 



t 



B 




Fig. 64. Broken conidiophores of Aspergillus niger, NRRL No. 326, X 700. A, 
Conidiophore showing a clean break suggesting a glass tube. B, Broken conidiophore 
showing fibrous wall structure. C, Portion of conidiophore crushed and further 
revealing the fibrous structure of the wall. 

the exceedingly abundant isolates having the approximate measurements 
of sterigmata and conidia noted in van Tieghem's description. The reader 
can assume, therefore, that in the opinion of the authors any possible 
description for the species itself would read essentially like that already 
given for the series on pages 216-219. 

Species Characterized by Large Primary Sterigmata and Small Conidia 

4 . phoenicis (Corda) Thorn, in The Aspergilli p. 175. 1926. In describing 
the black Aspergillus found upon dates, Patouillard and Delacroix (Bui. 
Soc. Myc. France 7: 118-120. 1891) compared their material to specimens 



ASPERGILLUS NIGER GROUP 223 

in the museum labeled "Ustilago phoenicis" and attributed to Corda, thus 
establishing the identity of the organism of Corda, which was not recogniz- 
able from any previous references. A. ustilago Beck (1892) is described 
with the same measurements. The measurements of sterigmata place 
A. phoenicis in the section of the group with primary sterigmata 50 to 60m 
in length; it was first described as Ustilago phoenicis by Corda (Icones 
Fung. IV., p. 9, pi. 3, fig. 26. 1840) and transferred by Patouillard and 
Delacroix, as noted above, to 8. phoenicis (Corda) Patouill. and Delacr. 
If we accept the use of "phoenicis" attributed to Corda as correct, this 
species becomes the type of the section of the black Aspergilli with primary 
sterigmata of intermediate length and conidia not over 4m in diameter 
(fig. 61 E and 62 B). This was cited by Thorn and Currie as A. phoenicis 
(Corda) Pat, and Delacr., and continued recognition of the species, to 
cover a group of strains occasionally encountered, appears warranted. 

Aspergillus pulverulentus (McAlpine) Thorn, in Jour. Agr. Res. 7:10-11. 

1916. 

Synonym: £. pulverulenta McAlpine, in Agr. Gaz. X. S. Wales (1896) 
7: 302. 1897. See also Thorn and Church, The Aspergilli, 
p. 179. 1926. 

McAlpine's data include: "White to dirty yellow mycelium;" heads 
155 to 340m in diameter, radiate with chains mostly separate; conidiophores 
erect, stiff, up to 7 mm. by 20m with walls up to 5m thick; vesicles 70 to 170m 
in diameter, globose or nearly so; primary sterigmata up to 144m long by 
8m, secondary 14 to 18m long; conidia globose, rough, about 4m in diameter. 
Colonies with these general characters have been studied in culture at 
least twice and maintained for long periods; one came from Spain, the 
other from Texas. A. strychni Lindau (Hedwigia Bd. 43, Rept, 5, p. 
306-7. 1904) is one of this series. 

Light Colored Forms 

Aspergillus niger mut. cinnamomcus (Schiemann) n. comb. 

Synonym: A. cinnamomeus Schiem., in Ztschr. Induktive Abstam. u. 
Vererbungslehre, Bd. 8, Heft 112, pp. 1-35, 16 fig., 2 pi. 
(1 col.) 1912. See also Thom and Church, The Aspergilli, 
p. 164. 1926. 

Colonies upon Czapek's solution agar at room temperature, rapidly 
growing and spreading, producing an aerial growth of conidiophores and 
heads reaching a pale cinnamon upon maturity. Reverse only slightly 



224 A MANUAL OF THE ASPERGILLI 

colored in the same shade. Conidial heads not crowded, globose. Conidio- 
phores smooth, thick-walled, with upper portion more or less brown, about 
1.5 mm. in length by 12 to 20m in diameter. Vesicles up to 40 to 50 m in 
diameter, crushing readily. Sterigmata in two series; primary about 
15 to 20m by 3 to 5m, sometimes larger, secondary about 8 by 2 to 3/jl. 
Conidia 3 to 4m, thin-walled, globose or subglobose, smooth or nearly so, 
almost colorless when viewed singly, pale yellowish to cinnamon in mass. 
Diagnosis based upon culture NRRL No. 348 (Thorn No. 3534b) re- 
ceived from . Schiemann as a mutation induced by introducing potassium 
bichromate into the culture medium. Approximately the same mutant 
appeared in Steinberg and Thorn's series of induced mutations (1939, 1940). 
Occasional cultures close to A. cinnamomeus have ben obtained from un- 
known sources in nature. 

Aspergillus niger mut. Schiemanni (Schiemann) n. comb. 

Synonyms: A. Schiemanni (Schiemann) Thom, Jour. Agr. Res. 7: 13. 
1916. 
A. fuscus Schiemann, in Ztschr. Induktive Abstam, u. 
Vererbungslehre, Bd. 8, Heft Y 2 , p. 1-35, 16 fig. 2 pi. 
(1 col.) 1912. 
Not A. fuscus Bonorden (Bot. Ztg. Jahr. 19: 202. 1861); 
Not S. fusca Bainier (Bui. Soc. Bot. France 27: 29, PI. 1, fig. 5. 
1880) 

Colonies upon Czapek's solution agar at room temperature, rapidly 
growing and spreading, developing a surface growth of conidiophores and 
heads forming a crowded fruiting area 2 to 3 mm. deep in slanted tubes, 
becoming a shade of brown near fawn color (Ridgway, PI. XL); reverse 
yellowish (fig. 61 D). Conidial heads large, fairly crowded. Conidiophores 
coarse, 2.5 mm. or more long by 15 to 25m wide. Vesicles up to 50 to 60m 
in diameter. Sterigmata in two series; primary, 15 to 40m by 4 to 6m, 
sometimes larger, secondary 7 to 8m by 2 to 3m- Conidia thin-walled, smooth 
except sometimes a trace of markings, 3.5 to 4.5 or 5m in diameter. 

Culture NRRL No. 361 (Thom No. 3534C) was received from Schiemann. 
Culture NRRL No. 362 was received from Biourge under the same name but 
shows a much deeper brown color (near Natal brown, Ridgway, PI. XL). 

The mutant, A. niger mut. Schiemanni, is distinguished from the parent 
type of A. niger by the color and smoothness of its spores. The name, 
A . fuscus Schiemann, is invalidated by the prior usage of the names A . fuscus 
by Bonorden and S. fusca by Bainier. 

Mutations of approximately this same type have been secured from 
cultures of strain NRRL No. 67 (PL VI C) by ultra-violet irradiation 



ASPERGILLUS NIGER GROUP 225 

(Raper, Coghill, and Hollaender) ; from cultures of other black Aspergilli 
through Cathode ray irradiation (Whelden 1939); by cultivation in the 
presence of various chemicals (Steinberg and Thom: 1939, 1940); and 
finally by the appearance in plate culture under normal conditions of a light- 
spored sector in an apparently typical culture of Aspergillus niger. See 
discussion Chapter VI. Two isolates labeled A. awamori Nakazawa 
which resemble this mutant in color have been received from Japan, one 
from Hanzawa, the other from Nakazawa. Both produce primary ster- 
igmata less than 20m in length but otherwise are close to Schiemann's 
mutant. 

Probable Synonyms 

Many species have been described by investigators working at different 
periods and at widely separated stations which obviously belong within 
the Aspergillus niger series as it is here considered. Some of these will be 
briefly noted since they were reported to present unique cultural or morpho- 
ological features, or since they account for interesting or important bio- 
chemical reactions. 

A. giganteus Mattlet (Ann. Soc. Beige Med. Trop. 6: 36. 1926) was described as a 
new species with conidiophores 2 to 6 mm. long; conidia 4 to 5m in diameter, rough, 
black-brown; but without other marks of separation. It would seem to have been a 
rather extreme variant in the two characters. 

A. atropurpureus Blochwitz (Ann. Mycol. 32(1/2) : 86. 1934.) Not A. atropurpu- 
reus Zimmermann (Centralb. f. Bakt. etc., 2 Abt., 8, No. 5/7, p. 218. 1902). Bloch- 
witz proposed to use the name A. atropurpureus for the purple-brown members of 
the A. niger group as he obtained them from the tropics. He gave conidial measure- 
ments as 2.5 to 3.5m in diameter, and added that chemical differences in the coloring 
substances warranted separation from the black, or blacker forms. 

A.ficuum (Reich) Hennings, in Hedwigia, 34, p. 86. 1895; and Reichardt, in 
Verhandl. K.K. Zoll. Bot. Gesell. Wien, 17: 335, 1867. Regarded as A. niger by 
Wehmer, in Centralb. f. Bakt. etc., 2 Abt., 18, No. 13/15, p. 394-395. 1907. See also 
Thom and Church, The Aspergilli, p. 174. 1926. 

Slight differences in morphology between this and A. niger v. Tieg. are reported by 
Hennings, but disregarded by Wehmer. Culture NRRL No. 364 (Thom. No. 142), 
received in 1909 from Westerdijk under this name, was reported by Thom and Currie 
(1916) as the most rapid producer of oxalic acid of all strains of A . niger tested. There 
is no morphological basis for separating this strain from A. niger, and it is distin- 
guished in culture only by the tendency, after many years in laboratory culture, to 
produce rather floccose colonies. 

A.batatae Saito, in Centralb. f. Bakt. etc., 2 Abt., 18, No. 1/3, p. 34. 1907. Saito's 
organism as described is close to A. niger: colonies reach brownish-black through 
yellow shades; conidiophores 2 to 4 mm. by 12 to 20m, smooth, thick-walled, brown in 
upper portion; vesicles 35 to 50m, globose; primary sterigmata 24 to 40m long by 8m 
at apex, secondary 10 by 3.2m; conidia globose, brown, finely roughened, 4 to 5m in 
diameter. A culture under this name from Formosa (NRRL No. 363) presented the 



226 A MANUAL OF THE ASPERGILLI 

comparatively large conidia described by Saito but possessed no other marks which 
would assist in identification. 

.4. iuteo-niger (Lutz) Thorn and Church, in The Aspergilli, p. 166. 1926. Syn.: 
S. luteo-nigra Lutz, in Bull. Soc. Bot. France 53: 48-52. 1907. Thorn and Church 
(1926) found one or more black Aspergilli in which the conidia appeared smooth in 
ordinary laboratory slide amounts in which they were treated with alcohol followed 
by lacto-phenol or other mounting fluids. Fragments floating in the mounting 
medium led to the test of such conidia in dry mounts, in oil, and in pure glycerine. 
Under these conditions, the conidia showed the typical marks of the A. niger series. 
Since smooth conidia were the sole definite contrast between A. Iuteo-niger Lutz and 
.4. niger v. Tiegh., the strains studied were accepted as Lutz' organism. Since that 
time, it has been shown that many black Aspergilli, ranging widely among the variant 
forms encountered, pass through a physiological stage in which the outer wall and 
color bars, when subjected to alcohol or other fluid causing active osmotic currents, 
break in pieces and float away leaving the conidia smooth and colorless, or only partly 
shaded toward the dark color of the group. It thus appears that A. Iuteo-niger may 
be dropped as failing to designate any definite group of strains and as failing to pre- 
sent any character of diagnostic importance. 

ASPERGILLUS CARBONARIUS SERIES 

Species Characterized by Conidia in Excess of 5. 0^— Sterigmata in Two 

Series 

Aspergillus atropurpureus Zimmermann, in Centralb. f. Bakt., etc., 2 Abt., 

8, Xo. 5/7, p. 218. 1902. 

Conidiophores hyaline or somewhat brownish in age, up to 800 by 16m 
to 20/x ; vesicles 60 to 80m in diameter, hyaline to brown; sterigmata, primary 
16 by 6m, secondary 3 to 4m by 1.5 to 2.0m; conidia globose, rough, with 
prominent warts, purplish-black, 6 to 10m in diameter. Isolated from 
Coffea liberica, in Java. Culture not studied by us. The species is possibly 
valid, and is presented as representative of occasional forms characterized 
by small sterigmata and large purple-black spores. 

Aspergillus fumaricus Wehmer, in Ber. Deut. Chem. Gesell. 51, Xo. 14: 

1663-1668, figs. 1-6. 1918. 

This species was named, but not fully described by Wehmer, and was 
not distributed by him. Its biochemical activity as a producer of fumaric 
acid is covered in the paper cited above, together with the admission that 
it belonged to the .4. niger group. Culture Xo. 4668.2 (C. Thorn) received 
from Xeuberg under this name presents a variant strain of the group which 
may be described as follows: Colonies producing a mass of yellow my- 
celium; conidiophores scantily and tardily produced up to 1, 2, or 3 mm. 
long, by 20 to 22m in diameter, smooth, bearing large radiate, yellow-brown 
heads. Vesicles up to 60 or even 100m in diameter, with walls thin, easily 
crushed. Sterigmata in two series, primary sterigmata up to 15m by 4 to 



ASPERGILLUS NIGER GROUP 227 

5m, secondary sterigmata rather coarse, some growing out into aborted 
hyphae. Conidia commonly 5m, occasionally 7 to 8m in diameter, with 
lengthwise color bars as in A. niger but paler in color. 

Culture No. 4668.2 (Thorn) is accepted as correctly named. By de- 
scription, A. fumaricus Wehmer differs little from A. atropurpureus Zimm. 
except for the production of yellow-brown rather than purple-black conidial 
heads. Differences in color definition and discrimination by different 
workers tend to leave both species in doubt. 

Aspergillus fonsccaeus n. sp. 

Synonym: S. fusca Bainier, in Bui. Soc. Bot. France 27: 29, PI. 1, fig. 5 ; 
1880. (Bainier's material grown upon moist bread in Tou- 
louse in 1880, was preserved in Roumeguere's Fungi Gallici 
Exsiccati Xo. 995) 

Bainier described S. fusca in terms closely parallel to van Tieghem's 
A. niger, except that the conidia were about double that species in size; 
Bainier reported these as rarely exceeding 9.4m in diameter, while our exam- 
ination of the exsiccati showed them to be mostly 5 to 8.5m in diameter 
and marked as in A. niger and A. earbonarius. Da Fonseca, in Rio de Ja- 
neiro, contributed a culture (Thorn Xo. 4707.878, XRRL Xo. 67, and strain 
Xo. 67 of Herrick, May, and associates) possessing approximately these 
spore measurements, which has retained its characteristic features for more 
than 20 years and which has proved industrially useful. We believe, 
therefore, that recognition of a species with sterigmata of intermediate 
length and conidia about double the dimensions of A. niger van Tieghem 
is warranted. The name A. fonsecaeus is proposed since the binomial 
A. fuscus had already been used for an Aspergillus by Bonorden in 1861 
(Bot. Ztg. Jahrg. 19: Xo. 29, p. 202) and has been used at least twice 
subsequent to this. The species would then include strains characterized 
by large, subglobose, coarsely roughened conidia ranging from 5.5 or 6.0m 
to 8.5 or 9.0m (fig. 63 C), including the strain "67" used by Herrick, May, 
AVells, Moyer, et al. of the Industrial Farm Products Research Division, 
U. S. Department of Agriculture, Arlington Farm, Virginia, for the pro- 
duction of citric and gluconic acids (see literature citations pp. 290-295). 
The following description is based primarily upon strain XRRL Xo. 67: 
Colonies upon Czapek's solution agar growing rapidly at temperatures 
from 24° to 30° C, attaining a diameter of 7 to 8 cm. in eight to ten days, 
consisting of a basal vegetative mycelium that is largely submerged and 
colorless, and abundant conidial structures commonly arranged in more or 
less conspicuous concentric zones; heads carbon black or brownish-black, 
imparting to the colony a like coloration ; reverse colorless in young colonies, 



228 A MANUAL OF THE ASPERGILLI 

commonly darkening in age and often becoming almost black after 2 to 3 
weeks. Conidial heads large, globose, radiate or with chains of conidia 
massed in an indefinite number of loose divergent columns, commonly 300 
to 500^ but often attaining a diameter up to 1 mm. Conidiophores varying 
in length from 1.5 to 3.5 mm. but averaging about 2.0 to 3.0 mm., mostly 
20 to 30m in diameter with walls 2.0 to 3.0m thick, smooth, often colored 
in dark shades in the region beneath the vesicle. Vesicles globose, fertile 
over the entire surface, commonly 50 to 75m in diameter (fig. 62 C), usually 
in brown shades, often quite dark. Sterigmata in two series, usually 
brown, often dark: primary sterigmata variable in different heads and in 
different cultures, ranging from 15 to 20m by 6 to 8m in some to 35 to 45m 
by 10 to 13m in others; secondary sterigmata ranging from 8 to 14m by 5 
to 6.5m but averaging about 9 to 10m by 5 to 6m. Conidia large (fig. 63 C), 
globose, conspicuously roughened with prominent color bars, ranging from 
5.5 to 8.5m- 

Since strain "67" appears in the industrial fermentation literature as 
Aspergillus niger and has been consistently distributed under this name 
over a period of several years, it is not our purpose here to challenge this 
designation, for this binomial is often used in a very general sense to cover 
any black member of this group. We do wish to emphasize, however, that 
this strain does not represent the common type of black Aspergillus usually 
isolated in routine examination of soil and moldy materials in general. 
The fact that it possesses large spores is of the greatest value in checking 
its purity and further commends it for use in industrial operations. This 
strain was originally received from Rio de Janeiro, and recently Dr. 
Dorival M. Cardoso of Sao Paulo, Brazil, has reported (personal communica- 
tion) that he has repeatedly isolated large-spored forms apparently closely 
related to it. It would, therefore, appear likely that this large-spored form 
represents a type of organism much more abundant in South America than 
in this country. 

Aspergillus pulchellus (Speg.) Thorn and Church, in The Aspergilli, p. 181. 

1926. 

Synonym: Aspergillopsis pulchellus Speggazzini, in Myc. Arg. V. in Ann. 
Mus. Nac. Buenos Aires Ser. 3, t. 13: 436. 1911. 

This species was described with colonies intensely black; conidiophores 1 to 2 mm. 
by 18 to 20m, with walls darkened; vesicles 50 to 60m in diameter; primary sterigmata 
10 by 30m; and conidia 8 to 10m in diameter and rough. The species appears to differ 
from van Tieghem's A. niger principally in its very large conidia, and it is extremely 
doubtful if it could be separated from A. fonsecaeus as described above. 

A. dipus of Ferdinandsen and Wing (Bot. Tids. 30: 220, fig. 6. 1910) represents 
another organism undoubtedly close to the forms under consideration. The presence 
of conspicuous foot cells, upon which character the species was based, is common to 
all members of the group, hence, is not a valid basis for separation. 



ASPERGILLUS NIGER GROUP 229 

Aspergillus carbonarius (Bainier) Thorn, in Jour. Agr. Res. 7: 12. 1916. 

Synonym : S. carbonaria Bainier, in Bui. Soc. Bot, France 27 : 27-28. 1880. 

Colonies grown upon Czapek's solution agar show vegetative mycelium 
white or with some yellow in submerged areas, broadly spreading, more or 
less zonate; sclerotia produced upon the surface of the substratum in old 
cultures; fruiting areas carbon-black. Conidiophores colorless below, 
yellow to yellow-brown toward the apex, 4 to 6 mm. or more in length 
and up to 25m hi diameter, with walls smooth, sometimes as much as 4m 
in thickness. Heads globose, varying in diameter up to 500m- Vesicles 
up to 90m in diameter, fertile over the entire surface, commonly with contents 
yellow-brown to black and in old heads f orming with the primary sterigmata 
a hard, brittle, carbonaceous mass. Sterigmata in two series, primary 
sometimes one septate, from 20 to 40m long in young or small heads, and 
up to 120m long in large heads by 5 to 13m in diameter at the apex, secondary, 
8 to 14m by 3 to 6m- Conidia at first smooth, becoming rough when ripe, 
5.5 to 10.5m in diameter. Colonies grow well upon all culture media used, 
with temperature optimum below 37° C. A culture from Dr. A. F. Blakeslee 
(NRRL No. 369, Thorn No. 4030.1) reproduces in detail the morphology 
recorded by Bainier. A. carbonarius has also been received in culture from 
the Gold Coast of Africa. 

The same morphology was also found in one of Dr. Farlow's specimens, 
S. acini-uvae Caballero (Bol. R. Soc. Esp. Hist. Nat, 28: 429. 1928). 
This was described as it appeared upon rotting grapes as follows : vesicles 
75 to 105m by 73 to 98m; primary sterigmata 59 to 80m by 13 to 22m and 
secondary 15 to 20m by 5 to 7m ; conidia globose, rough, 6 to 10m- Blochwitz 
in "Die Aspergillaceen" (Ann. Mycol. 27(3/4): 204, 222, and 232. 1929), 
with part of the type specimen before him, declares it to be A. carbonarius, 
apparently without cultivating it. Mosseray (LaCellule 43 : 222. 1934) 
places Caballero's organism in Aspergillus pulchellus. 

We cannot agree with Blochwitz (idem. p. 221, 222, fig. 12) in describing 
the vesicle in A. carbonarius as subglobose, and figuring the head as hemi- 
spherical. Bainier's original figures show the vesicle fertile to the very 
base. This is characteristic of the strains observed from various sources. 

ASPERGILLUS LUCHUENSIS SERIES 

Species Normally Showing One Series of Sterigmata, Occasionally Two. 

Aspergillus luchuensis Inui, in Jour. Col. Sci. Imp. Univ. Tokyo 15: 469, 

PI. 22, fig. 1-8. 1901. See also Usami, in Centralb. Bakt. 

etc., 2 Abt., 43, p. 250. 1915; The Aspergilli, 

p. 171. PI. IV. 1926. 

Colonies upon Czapek's solution agar spreading rapidly, producing 
abundant conidiophores and conidial heads which give a purple-black color 



230 A MANUAL OF THE ASPERGILLI 

to the whole colony, reverse in pale yellow shades. Conidial heads globose, 
up to 250 to 300m in diameter, splitting in age into short columns of spores. 
Conidiophores up to 1500m by 10m, smooth, yellow toward the vesicle. Ves- 
icles yellow, up to 40m in diameter, fertile over the entire surface. Ster- 
igmata mostly in one series, 6m or a little larger by 3m ; branched sterigmata 
occasionally appear. Conidia globose, 3.5 to 4m, roughened with spines 
rather than bars of coloring substances. 

Representatives of the species have been received from Japan (NRRL 
No. 356; Thorn No. 4291.3), from West Africa, from Bermuda, and from 
various points in the United States. While not as abundant as forms 
with double sterigmata, isolates with this kind of head are not uncommon. 

Inui also described A. perniciosus (Jour. Coll. Sci. Tokyo 15, p. 473, 
T. XXI, figs. 9-12. 1901) with color data closer to A. wentii than A. 
luchuensis. Its morphology, however, seems to belong here. It has not 
been rediscovered and discussed adequately in relation to either group. 
Culture No. 4707.757 (Thom), received from da Fonseca in Brazil, possibly 
represents this species: colonies in center at least transiently greenish, 
but without true green color; conidiophores not crowded, sinuous, apparently 
smooth when observed with low magnifications but with traces of pitting 
evident when examined with an oil-immersion objective, up to 1000m by 
10 to 15m; primary sterigmata 10 to 16m by 3 to 4m, secondary up to 8m by 
2 to 3m ; conidia 4 to 5m in diameter, with yellow markings in the form of 
loops and bars. 

A. luchuensis var. rubeolas Shih (Lingnan Sci. Jour. 15(3): 374. 1933) 
differs from the species by becoming chocolate brown rather than black. 
This would suggest careful comparison with A. japonicus Saito. 

Aspergillus japonicus Saito, in Bot. Mag. (Tokyo) 20: 61, 5 figs. 1906. 

Colonies upon Czapek's solution agar growing rapidly and spreading 
evenly, characterized by its purple-brown heads and the presence of few 
to many light brown sclerotia ; reverse colorless or nearly so. Conidiophores 
500 to 1000m by 12 to 15m, figured as showing concretions on the surface, 
with walls more or less brown. Vesicles globose, fertile over the whole 
surface, with walls brown and marked by the bases of sterigmata. Ster- 
igmata in one series, 7 to 9m by 5 to 6m, commonly falling away in mounts 
from old cultures. Conidia globose, echinulate, 4 to 5m in diameter. 
Sclerotia scattered throughout the colony, 650 to 1000m in diameter, white 
to pale yellow in color and often overgrown by mycelium and conidiophores. 

Type material has not been seen. The diagnosis is based upon two 
strains contributed by Dr. A. F. Blakeslee (NRRL No. 358: Thom No. 
4030.3, and NRRL No. 359: Thom No. 4030.5) which come fairly close to 



ASPERGILLUS NIGER GROUP 231 

Saito's description. A purple-brown strain (Thorn No. 5362.7) collected 
by Manns in Honduras showed slightly darker colors and slightly different 
measurements. A strain received from Raistrick as coming from Blochwitz 
through the Centraalbureau in Baarn labeled A. alropurpareus Zimmermann 
resembled the two Blakeslee cultures quite closely. It certainly did not 
comply with Zimmermann 's description (see p. 226). A. luchuensis var. 
rubeolis Shih is probably closely related if not identical with .4. japonicus 
Saito. 

Aspergillus violacco-fuscus Gasperini, in Atti. Soc. Toscana Sci. Nat. Pisa, 

Mem. 8, fasc. 2, p. 326. 1887. 

Colonies upon Czapek's solution agar, comparatively slow growing 
(fig. 61 F), purplish-brown with a faint violet shade, passing to purple drab 
in age, reverse colorless to dark purplish. Conidial heads purplish-brown, 
globose, not crowded, 100 to 150m in diameter. Conidiophores mostly 
less than 1 mm. in length but sometimes reaching 2 mm., 12 to 18m in di- 
ameter. Vesicles globose, varying up to 60m in diameter. Sterigmata 
generally in one series (fig. 62 D), 5 to 8m by 3m, occasionally in two series 
with secondaries 2 to 4m long. Conidia elliptical, 3.5 to 5m by 5 to 6.5m, at 
first hyaline, becoming violaceous, somewhat roughened. 

By description this is a variant member of the great group of black 
Aspergilli characterized by short sterigmata and elliptical conic|ia. 

Gasperini's material has not been seen but three cultures have been 
studied which are believed to differ from his species only in having some- 
what smaller conidia. The first of these was received in 1914, from Puerto 
Rico (NRRL No. 360: Thorn No. 3522.30), while the others were subse- 
quently obtained from Jamaica and from Professor Raistrick in England. 

In cultivating black Aspergilli an occasional strain produces heads at 
first showing a single series of sterigmata; then, as the colony becomes 
older, heads with both primary and secondary sterigmata dominate the 
culture. Upon careful examination many strains show both large heads 
with sterigmata in two series and, on shorter conidiophores mixed among 
the larger ones, small heads with simple sterigmata only or with both 
types mixed. Colonies of this kind probably accounted for .4. nanus 
Montagne, .4. subfuscus Johan-Olsen (Sopp.) and are known to account for 
A. pyri English. 

Aspergillus nanus Montagne, in Syllog. Generum Specierumque Cryptogamarum, 
p. 300, No. 112, Paris, 1856. Sacc. Syll. 4: 71. 1886. Species reported as a member 
of the black group with a single series of sterigmata about 15m in length and spores 
3m in diameter. This may have represented young fruiting structures of a typical 
A. niger. 

Aspergillus subfuscus Johan-Olsen, in Meddelelser fra Xaturh. forening i Kris- 



232 A MANUAL OF THE ASPERGILLI 

tiania, 1885. Described as showing smooth globose spores, 3.0 to 3.5m and a single 
series of sterigmata in culture, up to 20m in length, but with some secondary sterig- 
mata seen in the original material. The separation of this from A. niger appears 
questionable. 

Aspergillus pyri English n. n., in Doctoral Thesis, State College of Washington, 
Pullman, Wash. pp. 76-78, 1940; cited in abstract. A form described as showing a 
single series of sterigmata 14.4 to 19.2m by 3.6m and spinulose conidia 3.6 to 4.8m in 
diameter. In personal correspondence of June 1943, English stated that as a result 
of continued study of this strain and the finding of double sterigmata, he ques- 
tioned the desirability of maintaining the species designation. 

SYNOPSIS OF SPECIES PROPOSED BY MOSSERAY 

Biourge, who was a discriminating collector, accumulated 63 strains of 
"black" (or related) Aspergilli for each of which he could see sufficient 
individuality to warrant preservation. Mosseray, working in Biourge's 
laboratory, studied these strains, attempted to establish species lines among 
them, and proposed new specific names for all forms which he believed 
undescribed. With the whole 63 strains in parallel culture, all bearing the 
names applied by Mosseray, Biourge and Simonart were inclined to with- 
draw part of Mosseray's new species names, a position with which the 
writers heartily agree. Nevertheless, to present one concept of the range 
of variation confronted by the student of this group, Mosseray's synopsis 2 
has been translated with minor emendations, and is herewith presented. 

Mosseray's Synopsis 

A. Conidia 6 to 10m in diameter, rough; vesicles subglobose; primary sterigmata often 
100m or more in length; colonies jet-black. 

a. Sporulation more or less dense ; heads large ; reverse fumose or very dark olive 

with mycelium more or less wrinkled. 
Conidiophores 2, 4, or even 6 mm. long; sclerotia present in "natural" media 

A. carbonarius (Bainier) Thorn and Currie 

Conidiophores 1 to 2 mm. long; sclerotia not reported A. pulchellus (Speg.) 

Thorn and Church, Syn. S. acini-uvae Caballero 

b. Sporulation less dense; heads small, also with very small heads with single 



2 Translated and emended from Mosseray, Raoul, Les Aspergillus de la section 
"niger" Thorn and Church, in La Cellule XLIII: 271-273. 1934. All data are based 
upon colonies grown in slanted test tubes using Biourge's formula of "neutral Raulin 
agar" of the following composition: 

Water (distilled) 1000 cc . (NH 4 ) 2 HP0 4 0.400 g. 

Sucrose 50 g. (NH 4 ) 2 S0 4 0.200 g. 

Tartaric acid 0.40 g. FeS0 4 cryst 0.050 g. 

MgCC-3 0.250 g. ZnS0 4 0.050 g. 

NH 4 N0 3 2.500 g. Agar 20.0 g. 

K 2 C0 3 0.400 g. 

Sterilized at 120°C. for 20 minutes. 



ASPERGILLUS NIGER GROUP 233 

sterigmata and on very short conidiophores among them; mycelium gray- 
yellow; reverse dark reddish-brown and mycelium much wrinkled 

A. pseudo-carbonarius (Bainier nn.) Mosseray 

B. Conidia 2.5-4, even to 5m in diameter, mostly more or less rough and more or less 
colored but some smooth or nearly so ; vesicles normally globose ; primary sterig- 
mata mostly 20m long or longer, sometimes up to 100m. 
a. Colonies deep purple-brown or clear purple-brown. 
I. Conidiophores short (up to 2 mm. on an average). 

a. Wrinkles in reverse shallow, transverse (in tubes). 

Heads small ; conidiophores short (up to 1 mm.) 8 to 15m in diameter, 
primary sterigmata 5 to 20m; reverse cream colored 

A. microcephalus Mosseray 
Heads "medium"; conidiophores up to 2.5 mm.; primary sterigmata 
up to 45m; reverse dark reddish-brown, often spotted and becom- 
ing very dark A. phoenicis (Corda) Thorn 

Syn. A. longobasidia (Bainier n.n.) Mosseray 

Syn. A. bainieri Mosseray 1934 

/3. Wrinkles in reverse of mycelium fairly numerous, occasionally 

anastomosing. 

Appearance "mealy", deep purple-brown; conidiophores very short 

up to 0.5 mm. and 7 to 13m in diameter; primary sterigmata 4 to 

12m; reverse deep olive-brown A. pseudo-citricus Mosseray 

Appearance sub-granular; brown-purple, at times clear brown to 
umber; conidiophores up to 1.5 mm. by 7 to 20m; conidia smooth or 
nearly so; growth rapid. Reverse slightly colored: 
Reverse cream to pale brown; primary sterigmata 8 to 20m 

A . fuliginosus Peck 

Syn. S.fuliginosa Bainier 

Syn. A. praecox Mosseray 1934a 

Reverse very deep brown; primary sterigmata up to 50m; sclerotia 

occasional A. sclerotifer Mosseray 

Reverse uncolored to citron yellow; primary sterigmata 8 to 20m 

A. citrino-niger Mosseray 
7. Wrinkles sharp and numerous (reticulated). Reverse becoming 
rapidly dark, almost black. 

Aspect mealy; color purple-brown; conidiophores up to 1 mm; 
drops not colored; primary sterigmata 12 to 30m; conidia smooth 

or nearly so A . densus Mosseray 

Aspect mealy to granulate, deep purple-brown; drops numerous, 
bronze; primary sterigmata 12 to 30m. • -A. rutilans Mosseray 
Aspect subgranular, brown-purple; drops numerous, black, some 
of them very large; primary sterigmata 12 to 30m 

A. guttifer Mosseray 
Reverse not so dark, sometimes olive or colorless. 
Reverse deep olive, often spotted; primary sterigmata 15 to 30m 

A. Buntingii Mosseray 
Reverse not colored; commonly showing areas sterile or free 
from spores; mycelium slightly yellow at first; primary sterig- 
mata 20 to 50m A. variegatus Mosseray 



234 A MANUAL OF THE ASPERGILLI 

II. Conidiophores up to 3 mm., rarely 4 mm. long. 
a.. Sporulation normal. 

Reverse uncolored or slightly olive; drops few or none; primary 
sterigmata 20 to 40m; mycelium often yellow at first 

A. niger Van Tiegham 
Reverse orange-brown or purple-brown; drops large, black; pri- 
mary sterigmata 8 to 25m; mycelium colorless or rarely yellow 

A. Biourgei Mosseray 

Reverse pale reddish-brown; drops very few; heads very small; 

primary sterigmata 15 to 30m; mycelium colorless or sometimes 

rose; sporulation very slow A. Churchii Mosseray 

Reverse golden yellow; drops none; primary sterigmata 20 to 40m; 
mycelium reddish-yellow at first 

A. luteo-niger (Lutz) Thorn and Church 
Reverse dark olive-brown; conidiophores up to 1 to 2 mm.; primary 
sterigmata very variable; vesicles subglobose 

A. anomalus Mosseray 

Reverse cream to brown, spotted slightly, wrinkled with a dark 

band in center, sporulation scattered, pale, conidiophores up to 

4 mm. long A. tubingensis (Schober) Mosseray 

/3. Sporulation abnormal. 

Sporulation absent in patches and in the margin, with a granular 
appearance, and proliferation of mycelium throughout the co- 

nidial area A. granulatus Mosseray 

Sporulation massed at the thin end of the agar, less dense toward 

the bottom of the tube; mycelium wooly. 
Mycelium wooly, gray-white or yellowish-gray, carrying on the 
thin areas numerous simple heads ("fumigatiformes") and some 
normal heads. Reverse cream, slightly wrinkled 

A. velutinus Mosseray 
Mycelium less wooly, white; conidiophores 1 to 3 mm. long, very 
abundant at the thin end of the agar, the remainder sterile. Re- 
verse much wrinkled, cream. .... A . ficuum (Reich.) Hennings 
III. Conidiophores tall, up to 1 cm.; sporulation scanty, mostly toward the 
thin end of the agar. 

Heads large, splitting into divergent columns; conidiophores up to 
1 cm. by 30m; primary sterigmata up to 100m; reverse cream or slightly 

rose A . elalior Mosseray. 

Heads small, mycelium gray-rose; conidiophores rarely up to 1 cm., 
more often 2 to 6 mm.; very numerous little "fumigatiform" heads, on 
short conidiophores at the surface; reverse clear rose or salmon 

A. pseudo-elatior Mosseray 

IV. Conidiophores very irregular from less than 1 mm. to 3 mm. with much 

larger heads; reverse clear reddish-brown; mycelium yellow at first 

then dark brownish-red; sporulation delayed. A. pseudo-niger Mosseray 

b. Colonies not purple-brown but clear brown; morphology of A. niger; conidia 

mostly smooth or nearly so. 

I. Colonies clear brown or sepia; conidiophores up to 1 mm. ; reverse olive or 
lighter; with reticulated wrinkles; vesicles 20 to 50m; primary sterig- 
mata 12 to 50m; conidia smooth A. olivaceo-fuscus Mosseray 



ASPERGILLUS NIGER GROUP 235 

II. Colonies umber; conidiophores 1 to 3 mm.; reverse reticulated, dark 

reddish-brown; conidia smooth or nearly so A. Schiemanni Thorn 

Syn. A. fuscus Schiemann 
III. Colonies reddish salmon; conidiophores 1, 2, or even 3 mm.; reverse 
slightly wrinkled, uncolored, or pale cream; conidia smooth 

A. cinnamomeus Schiemann 

C. Conidia 3 to 5m, globose, smooth, slightly colored; vesicles globose; conidiophores 

up to 1 mm., slender ; primary sterigmata 12 to 20m; colonies appearing somewhat 
granular, deep brown; reverse olive passing to bronze; mycelium sulphur yellow 
at first A . citricus (Wehmer) Mosseray 

D. Conidia 3 to 5m, globose or elliptical, smooth or slightly rough ; colonies violaceous. 

a. Sterigmata in two series; vesicles globose; colonies purplish-violet or mauve; 

reverse violet-brown: conidia 3 to 5m, globose, smooth, uncolored 

A. awamori Usami 

b. Sterigmata in one series, short; vesicles subglobose; colonies in violaceous 

shades to mauve. 

Colonies mauve ("violet livide"), reverse uncolored, wrinkled; conidia 
globose or obovate, smooth 3 to 5m in long axis; sporulation slow; narrowly 
growing A . malvaceus Mosseray 

Colonies violaceous or dark violet slate; reverse dark yellow or orange, 
slightly wrinkled; conidia globose and rough, 3 to 4.5m; sporulation very 
rapid, and colonies broadly spreading, with sclerotia common on rice or 
other "natural" substrata, rare upon sugar media A.japonicus Saito 

Colonies violet-brown; reverse purplish-brown, wrinkled; conidia globose, 
rough, 3 to 5m in long axis; sporulation slow and more or less incompletely 
covering the surface; dwarf heads abundant. .A. atro-violaceus Mosseray 

Colonies dark brown or carob brown, with a mealy or granular appearance, 
reverse dark brown, or olive at the margins, wrinkled; conidia smooth, 
globose, 2.5 to 4m; vesicles globose or pyriform; primary sterigmata 3.5 to 
7.5m in diameter -4.. atro-fuscus Mosseray 

If the material available for study were limited to Biourge's 63 cultures, 
identification to strain, variety, or species might be possible. When Mos- 
seray subsequently returned to Brussels as Mycologist at the Jardin 
Botanique de l'Etat and was confronted by hundreds of other strains 
largely from the Belgian Congo, many of which presented further varia- 
tion, he began to see the impossibility of describing them all in terms which 
would permit subsequent identification. Biourge at that point (personal 
conference) withdrew his support from many of the diagnostic features 
accepted in Mosseray's memoire and agreed to the proposal that specific 
names in black Aspergilli could only be serviceable as bringing together 
aggregates of strains showing common and fairly dependable morphological 
characters. This was the attitude held by Thorn and Church in The 
Aspergilli (1926) based upon the examination of many hundreds of black 
Aspergilli, and it remains the position of the writers at this time. 

Since variation is characteristic of the whole genus — not one group 



236 A MANUAL OF THE ASPERGILLI 

alone, the subject is covered in the chapters on Morphology and on Varia- 
tion. 

Coloration 

Color has been emphasized in most of our attempts to separate these 
strains in culture. Linossier (1891) extracted from his strain of A. niger 
a coloring substance soluble in hot water which he called aspergilline. 
This substance is readily demonstrated. Blochwitz (1929, p. 219) reports 
this material to be soluble in "NH 3 and alkalies," and we have extracted it 
with hot water. Microscopic comparison of conidiophores, heads, and 
separate conidia from numerous strains leads to the conclusion that the 
same substance may give a brown color to the upper one-third of the 
conidiophore, to the vesicle and its contents, to the walls of the sterigmata, 
and in A. carbonarius so fill the whole vesicle and sterigmatic area with a 
brittle mass as to justify Bainier in calling the head carbonaceous. Conidia 
in a few strains have appeared to possess smooth, uniformly brown cell 
walls. As a group, however, the black aspergilli have globose conidia with 
a firm, uncolored or slightly colored inner wall, a very thin outer wall, and 
between the two the coloring substance, presumably aspergilline, deposited 
as granules, warts, or bars running lengthwise of the cells and presenting 
a pattern characteristic of the whole group. When pale-colored variants 
(mutants?) such as A. cinnamomeus (p. 223), or darker ones such as A. 
schiemanni (p. 224), or the variously brown to deep carbon black ones 
like A. carbonarius are compared, the relative quantities of coloring matter 
seem to account for the progressive darkening of the head colors as de- 
scribed in the series. 

In the "reverse", or underside, of the mycelium and in the culture 
substratum, a range of shades from colorless to yellows to reddish-brown, 
and even very dark shades, is reported for particular species growing upon 
specified substrata. The chemical reactions back of these colors are not 
known for the black aspergilli. Blochwitz has reported extractions of 
color from various species of Aspergillus and other molds, and observed 
that variations in these colors were readily obtained with reagents. Some 
of the extracted materials were reported to act as indicators. It is, there- 
fore, doubtful whether the succession of colors present in such a related 
series of organisms represent substances differing .fundamentally one from 
another, or merely successive steps in the transformation and reactions of 
the same general type of product. 

Occurrence and Economic Importance 

The black aspergilli are probably more common than any other repre- 
sentatives of the genus. They are world-wide in distribution and occur 



ASPERGILLUS NIGER GROUP 237 

in and upon the greatest variety of substrata, including grains, forage 
products, spoiled fruits and vegetables, exposed cotton textiles and fabrics, 
leather, dairy products and other protein-rich substrata, and decaying 
vegetation in the field. They are abundant in all soils examined; and from 
studies which have been made by the authors and other investigators, it 
would appear that they are particularly abundant in soils from tropical and 
sub-tropical areas. 

With the possible exception of the A. flavus-oryzae group, which is of 
great economic importance in the Orient, the black aspergilli are un- 
doubtedly more widely used in industry than any other group of molds. 
Since the present volume is primarily a manual designed to assist the 
worker in the study, diagnosis, and maintenance of the aspergilli that come 
into his hands, no attempt will be made to discuss the various fermentations 
and other biochemical activities of the black aspergilli. However, these 
fermentations are of great importance and will be briefly noted. For the 
reader who is interested in these fermentations, or perchance, is actually 
conducting them, a fairly complete list of references is presented for each 
in the "Topical Bibliography" (Chapter XXII). In each case it has been 
our aim to present sufficient references to provide the reader with a reason- 
ably comprehensive guide to the literature of the field. 

Gallic Acid: Raulin and his coworkers in Paris during the early 1860's 
identified the organism active in the production of gallic acid by the fermenta- 
tion of gallnuts and other tannin-bearing substances. The species was 
first discussed as Ascophora nigrans. Van Tieghem in 1867, named and 
described the organism correctly as A. niger. Recurrent investigations 
have been conducted on this fermentation from that period to the present 
time (see Topical Bibliography, p. 294). 

Citric Acid: In 1917 Currie published his fundamental studies on the 
formation of citric acid by strains of A. niger and thereby established the 
basis for one of the most important of all industrial mold fermentations. 
Subsequent to this, investigators in the United States and abroad have 
made many additional and important contributions. While no attempt 
will be made to cite all of these, the works of Bernhauer, Wehmer, Doelger 
and Prescott, and the U. S. Department of Agriculture group, including 
Wells, May, Moyer, Herrick, and Ward, are considered to be outstanding. 
A selected list of references to the citric acid fermentation is presented 
on pages 290-293. 

Fumaric Acid: Certain strains of the A. niger group produce appreciable 
amounts of fumaric acid, and it was to one of these forms that Wehmer in 
1918 applied the n&me A. fumaricus. At the present time, however, fumaric 
acid is produced in industry by fermentation with species of Rhizopus 
rather than strains of the black aspergilli (see p. 293). 



238 A MANUAL OF THE ASPERGILLI 

Gluconic Acid: Selected strains of A. niger are used industrially for the 
production of gluconic acid. This process, like the citric fermentation, 
has been investigated by many workers. Outstanding contributions have 
been made by Molliard, Bernhauer, and the U. S. Department of Agri- 
culture group, including Herrick, May, Wells, Moyer, Gastrock, Porges, 
and others (see pp. 295-297). 

Oxalic Acid: Under certain conditions some strains of A. niger produce 
appreciable quantities of oxalic acid. While it is usually avoided rather 
than encouraged, this fermentation has been investigated by Wehmer (1891 
and 1892), Raistrick and Clark (1919), Jacquot (1938), and others (see 
pp. 298-299). 

The production of these various acids in quantities sufficient to have 
economic importance represents to an appreciable degree specific strain 
characteristics, and the greatest possible care must be exercised in main- 
taining these strains in a state of high productivity. It has been found, 
however, that in certain cases the same strains can be made to produce 
substantial yields of two or more of these acids by varying the composition 
of the nutrient solution and certain environmental factors. The reader is 
referred to the extensive literature on this subject. 

Enzymes: While they are not commonly cultivated for the production 
of enzymes as such, as are members of the A. flavus-oryzae group, the black 
aspergilli produce a number of enzymes in appreciable quantities. Be- 
ginning with Fernbach in Germany (1890) and Bourquolet (1893) in 
France, various authors have devoted considerable study to their formation. 
A number of papers relative to enzyme production by members of the group 
are cited in the "Topical Bibliography" under the subtitle "Enzymes of 
Aspergillus niger" (pp. 294-295). 

Fat Production: The mycelium of A. niger contains appreciable fatty 
materials (Pontillon, 1932; Bernhauer and Patzelt, 1935; Schmidt, 1935; 
and others). The waste mycelium from the citric acid fermentation is 
reported to provide a satisfactory source of sterols for irradiation in the 
production of Vitamin D. 

Soil Testing: A. niger has been successfully employed as an assay 
organism for determining mineral deficiency of soils, particularly de- 
ficiencies in phosphorus and potassium. Papers on the so-called A. niger 
method of soil testing were first published by Kiessling and Schmidt and 
by Schlots, Smith, and Brown in the same year (1932), to be followed by 
more elaborate studies by Stock (1933), Niklas and associates (1933), and 
others. Additional references to this method are presented in the "Topical 
Bibliography" (pp. 313-314). The strain employed by Niklas is main- 
tained in the culture collection of the Northern Regional Research Lab- 
oratory as No. 323. 



ASPERGILLUS NIGER GROUP 239 

Mildew: Strains of A. niger represent a common cause of mildew on ex- 
posed wood surfaces and cotton fabrics. Partansky and McPherson 
(1940) used a strain of this species successfully for testing the mold- 
resistant properties of oil paints. Aspergillus niger is commonly included 
in the mixtures of miscellaneous molds used for testing the effectiveness 
of mildew- and rot-proofing agents when impregnated in textiles and fabrics. 
Where pure cultures are employed, species of Chaetomium and Metarrhizium 
are generally used. 

Mold Physiology 

Members of the Aspergillus niger group have been used extensively in 
investigations on mold physiology, probably more than any other form. 
As early as 1909 Latham studied nitrogen assimilation by A. niger (S. 
nigra) to be followed in 1911 by Dox studying phosphorus assimilation by 
the same species. Beginning in 1918 and continuing up to the present 
time, Steinberg has published a succession of papers on the physiology of A . 
niger, with special reference to the role of heavy metals in its nutrition. 
A single strain of A. niger which is carried in the NRRL collection as No. 334 
(Thorn No. 4247) has been used throughout these investigations. Studies 
of a somewhat similar character have been conducted by Bortels (1927), 
Levy (1932), Gollmick (1936), and others. Citation of these papers, to- 
gether with many additional references are presented in the Topical 
Bibliography under the subtitle "Physiology". An attempt has been 
made to present sufficient references to serve as a point of entrance to the 
literature of the field. 

Pathogenesis 

Members of the Aspergillus niger group are commonly isolated from 
the external ear of man, this being the source of the classic Sterigmatocystis 
antacustica of Cramer. Other species reported to have been isolated from 
cases of otomycosis include A. niger van Tieghem, A. phoenicis (Cda.) Thorn 
and Church (with long primary sterigmata), A. giganteus (Mattlet) Dodge, 
and A. Macfiei Dodge. A. Macfiei showed no signs of pathogenicity 
when tested on experimental animals. There is no morphology to dis- 
tinguish either of the latter two forms from the ubiquitous saprophytic 
types, and it appears probable that many strains can become established 
in the auditory canal under certain favorable and probably temporary 
conditions. 

Once entrenched in the flesh about the auditory canal, the mycelium has 
been found to be very persistent. One case is known in which occasional 
abscesses have occurred over a period of 25 years during which desultory 
treatment has quieted the inflammation but has not destroyed the parasite. 



240 A MANUAL OF THE ASPERGILLI 

In the same way, extensive development of these forms as points of 
infection in the lungs may give rise to conditions diagnosed as pseudo- 
tuberculosis which may persist for long periods without resulting either in 
death or complete recovery of the patient. Spores of A. niger are air 
borne and hence are commonly drawn into the respiratory tract. They are 
occasionally reported as causative agents in allergic reactions. 

Single Strain Cultures 

The great importance of some of the black aspergilli as fermentative 
agents makes punctilious preservation of the actual strain employed the 
only means either of insuring a process against breakdown or of intro- 
ducing it in a new locality. Even when such a culture is maintained with 
the utmost care, natural variation sometimes occurs. Induced variation, 
such as that described by Steinberg and Thom in a series of papers (1939-40) 
presents a further hazard in processes where cultures are grown at extreme 
H-ion concentrations, or in the presence of chemical or other substances 
which may actually be toxic at the concentrations employed. In a process 
in one laboratory a strain appeared that presented a colony aspect in which 
only a few typical black heads developed in a background of dwarfed and 
fractional conidial fruiting structures. In another case, a mutant appeared 
which differed so radically from A. niger in its secondary sterigmata and 
spore chains that the Yuills described it as a new genus and species, Clado- 
sarum olivaceum (Trans. Brit. Myc. Soc. 22: 194-200, Pis. 11-13. 1938). 

The user of one of these Aspergilli, then, needs to know his organism in 
cultural aspect, in microscopic details, and in essential reactions in standard- 
ized and reproducible substrata. Furthermore, he must be able to main- 
tain it free from contamination with other species, and likewise free from 
such variation, either natural or induced, as will interfere with consistent 
and controlled results. 



Chapter XVIII 
THE ASPERGILLUS WENTII GROUP 

Outstanding Characters 

Conidial heads large, typically globose, often splitting irregularly in 
age — varying in color from dull yellowish to ecru-olive, and from light 
to dark brown depending upon the species and strain. 

Conidiophores smooth-walled or nearly so, but often appearing finely 
roughened when examined in dry mounts. 

Vesicles globose, fertile over the entire surface. Sterigmata in two 
series. 

Conidia commonly elliptical, smooth or somewhat roughened depending 
upon the species. 

Sclerotia present or lacking, dark brown to black, characteristically 
white-tipped when young. 

The Aspergillus wentii group as presented here is recognized as somewhat 
artificial, in comparison with such strictly natural groupings as the A. 
glaucus, A. nidulans, and A. clavatus groups. The degree of relationship 
between the species included is open to question. Yet all of the forms 
possess certain characteristics in common: (1) conidiophores are smooth- 
walled, or nearly so; (2) conidial heads are large and strictly globose, at 
least when young; and (3) all appear to occupy taxonomic positions some- 
what intermediate between the Aspergillus niger group on the one hand 
and the Aspergillus flavus or Aspergillus ochraceus groups on the other. 

Group Key 

I. Conidia smooth-walled. 

A. Sclerotia lacking; vegetative mycelium and young conidiophores reddish in 

color. 

1. Conidial heads light brown, near wood brown (Ridgway PI. XL) 

A. panamensis Raper and Thorn 

B. Sclerotia present, dark brown to black, vegetative mycelium colorless, and 

young conidiophores colorless or pale yellow. 

1. Conidial heads dull yellow to ochraceous; sclerotia globose or nearly so 

A. alliaceus Thorn and Church 

2. Conidial heads in yellow-green shades near ecru-olive (Ridgway PL 

XXX) A. avenaceus G. Smith 

II. Conidia more or less echinulate. 

A. Sclerotia present or lacking, depending upon the strain and the substratum; 
conidial areas in orange-brown to brown colors. 

1. Conidiophores colorless; colonies often conspicuously floccose 

A . wentii Wehmer 

2. Conidiophores brown (See A. niger) A. hennebergi Blochwitz 

241 



242 A MANUAL OF THE ASPERGILLI 

Aspergillus panamensis Raper and Thorn, in Mycologia 36: 568-572, 

fig. 5. 1944. 

Colonies on Czapek's solution agar at room temperature very thin, 
consisting of a sparse and transparent growth of vegetative hyphae, almost 
wholly submerged, bearing widely scattered, erect conidial structures 
with radiate heads, light brown in color. Colonies upon malt extract agar 
at room temperature growing well and fruiting luxuriantly, reddish brown 
in color, consisting of a dense basal mycelium, predominantly red, from which 
develop massed condial structures in broken or continuous concentric 
zones (fig. 65 A), many conidiophores abortive and sterile, fertile co- 
nidiophores bearing globose to radiate heads, light brown in color, near wood 
brown (Ridgway, PI. XL) ; these, together with red-colored sterile structures 
and aerial hyphae, give the colony its characteristic appearance and color; 
in age, colonies tending to develop a loose floccose overgrowth, more or 
less obscuring the abundant conidial heads; reverse dull brown; odor none. 

Conidial structures arising directly from the substratum, scattered or 
abundant, depending upon the culture medium employed (fig. 65 B). 
Heads typically globose, in age characterized by loosely radiating chains 
of conidia, less commonly by few to several roughly columnar masses variable 
in size, commonly ranging from 250 to 450m in diameter, occasionally up 
to 500m, varying in color from avellaneous to wood brown (Ridgway, PL XL) 
to Saccardo's umber (Ridgway, PI. XXIX). Conidiophores straight, 
mostly 600 to 900m in length by 9 to 12m in diameter, occasionally larger, 
with walls smooth, comparatively heavy, ranging from 3 to 3.5m thick in 
the basal area to 1.5 to 2m in the terminal area, approximately uniform in 
diameter throughout except for a limited reduction immediately beneath 
the vesicle. Vesicle colorless, comparatively thin-walled, globose or 
slightly elongate, mostly 25 to 30m in diameter, fertile over the entire area 
(fig. 65 C). Sterigmata in two series, closely packed, primaries 5.5. to 
6.5m by 2.4 to 2.8m, secondaries 5 to 6m by 1.5 to 2m. Conidia light yellowish- 
brown in mass, globose to subglobose, smooth-wal'ed, mostly 2.2 to 2.6m 
in diameter, occasionally 2.8m- 

Type culture NRRL No. 1785 was isolated in January 1942, from 
Panama soil collected by Mr. John T. Bonner. A second culture, NRRL 
No. 1786, differs from the above strain in minor details but clearly belongs 
with it. This was isolated from a second sample of Panama soil collected 
by Bonner. 

The species is considered to represent a form somewhat intermediate 
between the Aspergillus nigcr group and A. wentii. Superficially, at 
least, there is evidence of relationship with Aspergillus niger mut. cin- 
namomeus (syn. Aspergillus cinnamomeus Schiemann) and Aspergillus 
niger mut. Schiemanni (syn. A. fuscus Schiemann). It bears a certain 




Fig. 65. A-C, Aspergillus panamensis NIHIL No. 1785: A, Colonies upon malt 
extract agar, 10 days; B, Conidial heads, showing tendency to split into loose diver- 
gent columns, X 9; C, Conidial heads showing globose vesicles and sterigmata in 
two series, X 500. D-F, Aspergillus avenaceus NRRL No. 517: D, Single colony 
on Czapek's solution agar snowing numerous large sclerotia; E, Portion of above 
colony showing greater detail of heads and sclerotia, X 6; F, conidial heads showing 
large globose vesicles, X 160. 



243 



244 A MANUAL OF THE ASPERGILLI 

resemblance to these forms in the comparatively light color of its conidial 
heads and in the smallness and general character of its conidia, but differs 
from these forms in three very striking particulars. (1) It character- 
istically develops an extensive red-colored aerial mycelium upon media 
such as malt-extract agar where it attains its maximum growth; (2) it grows 
very sparsely upon Czapek's solution agar, upon which the above noted 
forms grow luxuriantly; and (3) it possesses very small primary sterigmata, 
measuring 5.5 to 6.5m by 2.4 to 2.8m in contrast to 13 to 15m by 3 to 5m for 
mut. cinnamomeus and 15 to 40m by 4.6m for mut. Schiemanni. Whether 
or not the species actually represents a naturally occuring mutation from 
A. niger can only be guessed. The smallness of its conidia and primary 
sterigmata would hardly support this hypothesis. In cases where mu- 
tations have been obtained from known cultures, the dimension of specific 
structures in such mutations generally agree very closely with those of the 
same structures in the parent strain; and black Aspergilli with the dimen- 
sions of A. panamensis are rarely, if ever, encountered in nature. The 
possibility of this representing a mutation is not excluded, but until ad- 
ditional evidence supporting such origin is forthcoming, the writers feel 
warranted in maintaining as a distinct species this unique form which 
obviously is able to maintain itself in the soils of Panama. 

The correct taxonomic position of this species remains in doubt. It is 
included with Aspergillus wentii, although we realize that this placement 
is not entirely satisfactory. As continued isolations are made from tropical 
soils and other sources it is our hope that additional forms may be found, 
which will furnish evidence of a more exact relationship. 

The very sparse development of A. panamensis upon Czapek's solution 
agar, containing sucrose, results from an invertase deficiency; when dextrose 
is substituted as a carbon source the fungus grows luxuriantly and fruits 
abundantly. 

Aspergillus alliaceus Thom and Church, in The Aspergilli, p. 163. 1926. 
Discussed without name as Thom No. 4660 by Walker and Lindegren, 
in Jour. Agr. Res. 29: 507-514. 1924; and by Walker, Lindegren, and 
Bachmann, in Jour. Agr. Res. 30: 175-187. 1925. 

Colonies on Czapek's solution agar rapidly and broadly spreading, with 
loosely floccose aerial sterile mycelium, bearing scattered ochraceous 
heads among abundant dark to almost black sclerotia (PI. VI D and fig. 66 
A) in some strains, predominantly floccose with limited conidial heads and 
few sclerotia in others (fig. 66 B); reverse uncolored. Conidial heads dull 
yellow to ochraceous, strictly globose when young and remaining radiate 
or splitting irregularly in age (fig. 66 D), up to 300m in diameter, often more 
abundant in cultures after many transfers. Conidiophores up to 150m 



THE ASPERGILLUS WENTII GROUP 



245 



long by 15m, with walls up to 1.5m in thickness, appearing smooth in liquid 
mounts, but showing rudimentary markings or pits when examined dry. 
Vesicles globose to subglobose, up to 40 to 50m in diameter, with walls 
about 2m in thickness, and showing pores where sterigmata are attached. 
Sterigmata in two series, primary 7 to 9m or even 12m by 3 to 4m, secondary 




A 



B 




Fig. 66. Aspergillus alliaceus. A, Heavy sclerotium-producing strain, NRRL 




(strain NRRL No. 318) showing tendency to split into divergent columns, X 18 



7 to 8m by 2m- Conidia elliptical to globose, yellowish, 2.5 or 3m in diameter. 
Perithecia not found. Sclerotia often very abundant (fig. 66 C) at first 
white but quickly becoming black or nearly so, in many strains dominating 
the character of the colony. 

Strains compared include NRRL Xo. 315 (Thorn No. 4656) from a dead 



246 A MANUAL OF THE ASPERGILLI 

blister beetle (Macrobasis albida), NRRL Xo. 316 (Thorn No. 4660) 
isolated from onion bulbs, and others with characteristic heads and scle- 
rotia have been obtained from garlic bulbs, from cactus plants, and from 
soils, particularly from the general region of Texas, Arizona, and Mexico. 
Strains have also been isolated from soils collected near Calcutta. 

The large, globose, pale yellow to ochraceous heads of the species strongly 
suggest relationship to the A. oehraceus group. The smooth-walled co- 
nidiophores and black sclerotia, however, more closely ally it with Asper- 
gillus wentii and the other species grouped with it. It may, however 
represent a form somewhat transitional between the great groups repre- 
sented by A. niger on the one hand and A. oehraceus on the other. 

Aspergillus avenaceus Geo. Smith, in Brit. Mycol. Soc. Trans. 25: 24-27, 

PL 1, figs. 1-3. 1943. 

Colonies on Czapek's solution agar (with sucrose) at room temperature 
spreading rapidly, more or less conspicuously zonate (fig. 65 D), slightly 
fioccose, white at first, then dull yellow to ecru-olive (Ridgway, PL XXX) 
as shown in PL VI E, with, at times, a greenish tinge without becoming 
truly green; reverse pale dirty pink. Conidial heads large, globose 400 
to 600m in diameter, or up to 1,000m, splitting into columnar masses of 
conidial chains. Conidiophores up to 5 mm. long, 18 to 30m in diameter, 
with walls 2.5 to 4m thick, smooth in fluid mounts; but appearing finely 
roughened when examined dry. Vesicles globose or slightly flattened, 
thick-walled, up to 185m in long axis, sterigmata in two series, primary 22 
to 50m by 6m, secondary 11 to 13m by 4m (fig. 65 F). Conidia ellipsoid, 
smooth, 4 to 6 or 6.5m by 3.2 to 4m- Sclerotia dark grayish-brown to 
black (fig. 65 E), elongate, irregularly flask-shaped, sometimes with the 
"neck" forked, apical portion white to gray during development, 2 to 3 mm. 
in long axis, scattered in concentric zones after 7 to 10 days. 

On Czapek agar with glucose, sclerotia are more abundant and larger. 
On wort, or potato agar, conidial heads are abundantly produced but 
sclerotia are delayed for several weeks and are few in number. 

Species characterization adapted from George Smith's description. 

This very distinctive species (NRRL 517: Thom 5725) is represented by 
a single isolation from seed peas made in 1938 by Dr. G. E. Turfitt of the 
London School of Hygiene and Tropical Medicine, University of London. 

Aspergillus wentii Wehmer, in Centralbl. f. Bakt. etc., 2 Abt., 2, p. 150. 
1896. See also The Aspergilli, Thom and Church, p. 183. 1926. 

Colonies on Czapek's solution agar, rapidly growing and broadly spread- 
ing, fioccose with white or yellowish aerial hyphae which in some strains 
pile up in the plate (PL VI F, and fig. 67 A) or fill the test tubes for several 



THE ASPERGILLUS WENTI1 GROUP 247 

centimeters (fig. 67 C), but remain inconspicuous in other strains (fig. 67 B) ; 
with developing heads at first white through yellow shades to olive-brown, 
medal bronze or snuff brown (Ridgway, Pis. IV, XVI, XXX, column 19, 
and XXIX, column 15 K.), or, according to Wehmer, coffee brown to 
chocolate brown ; reverse becoming reddish-brown in old cultures. Conidial 
heads large, globose, generally remaining radiate in age (fig. 67 D), ranging 
up to 500/x in diameter, changing from yellow shades to brown. Conidio- 
phores up to several millimeters in height by 10 to 25m in diameter, with 
walls colorless up to 4m in thickness, studded with droplets in growing 
colonies and often appearing slightly roughened when examined dry, 
but uniformly smooth in fluid mounts. Vesicles globose or nearly so 
(fig. 67 E), varying up to 80m in diameter, fertile over the entire surface. 
Sterigmata usually in two series, primaries 10 to 20m by 3 to 5m, occasionally 
much larger, secondaries 6 to 8m by 3/jl. Conidia borne in long chains, 
more or less elliptical, ranging from 3.5 to 6m in long axis, but mostly 4 
to 5m, double wall clearly evident, ranging from almost smooth to marked 
by ridges sometimes suggestive of A. niger, again more closely resembling 
the A. flavus series. No perithecia reported. Sclerotia often encountered 
(fig. 67 F), dark brown to black, ovate with long axis vertical. 

Culture description based upon strain NRRL No. 375 (Thorn No. 116) 
obtained in 1909 from the Centraalbureau as Wehmer's original organism, 
as well as numerous isolations from soils and other materials collected by 
the authors in the United States, and other strains contributed by in- 
vestigators from all over the world. 

Numerous strains with the general aspect of Wehmer's species have been 
seen from Java, China, South America, Japan, the Straits Settlements, 
British Guiana, and Brazil. In our experience it has been isolated from 
cottonseed cake, from olives, from soil, and from numerous other sources. 
It is to be regarded as very widely distributed and to be common on many 
types of decaying vegetable products. 

The variations in colony aspect in different strains run from an extreme 
of mycelial growth filling the test tube that is characteristic of cultures 
such as the Wehmer organism, to colonies forming a crowded surface 
growth of conidiophores only and distinguishable from .4. tamarii only by 
a lack of greenish color in the early fruiting period and in the characteristic 
smooth conidiophores and finely roughened conidia. 

Aspergillus archaeoflavus Blochwitz (Ann. Mycol. 31(1/2) : 73-83. 1933) represents 
a non-floccose form which is hardly separable from A. wentii. In our examination 
of the type strain (XRRL No. 382: Thorn Xo. 5346), received in 1933 from Baarn, 
measurements were somewhat less than those cited by the author. The absence of 
conidial markings, to which Blochwitz called attention, would not bar it from A. 
wentii since in some strains conidia are almost entirely smooth, in others finely rough- 
ened, while in still others they are conspicuously echinulate. 



248 



A MANUAL OF THE ASPERGILLI 











- 






A 



B 




Fig. 67. Aspergillus wentii. A and B, colonies of strains NRRL No. 375 and No 
385, respectively, upon Czapek's solution agar, 10 days. C, Tube cultures of four 
representative strains. D, Conidial heads from old culture, X 18 E Single head 
showing smooth conidiophore, globose vesicle, and sterigmata in two series, X 500. 
t , Detail ! of colony margin in strain NRRL No. 379 showing sclerotia and abundant 
conidial heads, X 9. 




Plate VII 

.4 (upper left), Aspergillus terricola var. americana Marchal, NRRL No. 424. B (upper right), Aspergillus 
tamarii Kita, NRRL No. 427. C (center left), Aspergillus oryzae (Ahb.) Cohn, NRRL No. 458. D (center 
right), Aspergillus flatus Link, NRRL No. 1957. E (lower left), Aspergillus quercinus (Bain.) Thorn and 
Church, NRRL No. 394. F (lower right), Aspergillus ochraceus Wilhelm, NRRL No. 39s. All cultures 
growing upon Czapek's solution agar. (Color photographs by Haines, Northern Regional Research Labora- 
tory. Reproduced through co-operation of Chas. Pfizer & Co., Inc.) 



THE ASPERGILLUS WENTII GROUP 249 

Aspergillus wetitii var. minimus Xakazawa, Takeda, Okada, and Si mo (Jour. 
Agr. Chem. Soc. Japan 10(2) : 176-177. 1934) shows measurements varying somewhat 
from those of the species and from those given by Blochwitz, but it is not sufficiently 
marked to warrant separation. 

Sterigmatocystis aerect Bainier (Bull. Soc. Bot. France 27: 28. 1881) may have been 
a member of this series but was not sufficiently described to permit positive 
identification. 

Aspergillus hennebergi Blochwitz, in Ann. Mycol. 33: 23S-239. 1935. The species 
is described as having the aspect and colors of a non-floccose A. wentii or an A. tamarii 
with red sclerotia but with conidiophores browned as in the partially browned con- 
idiophores of the .4. niger group. 

An albino "variant" of A. wentii was isolated by Mosseray (Ann. Soc. Sci. Brux. 
54: 161-189. 1934) from a normal culture of this species, and was found to retain its 
distinctive characters through repeated transfers in laboratory culture. No name 
was given to this mutant. 

Occurrence and Economic Importance 

Aspergillus wentii is a cosmopolitan species that is fairly common in 
soils, upon moist grains and other vegetable matter undergoing slow 
decomposition, and may be isolated less frequently from a wide variety 
of other materials collected from nature. It is apparently world-wide 
in distribution. In the Orient, it is often included with Aspergillus tamarii, 
A. flavus, and A. oryzae, all under the latter name as a rule, in the "Koji" 
preparations used in the manufacture of various soy products. Likewise, 
it has been investigated with these same species in connection with the 
production of various mold enzymes. At the same time, it has been 
included with the black Aspergilli in studies on the production of organic 
acids by molds. Recently Karow (1942) has reported one strain of this 
species to give substantial yields of citric acid in submerged culture. 
Yabuta (1912) reports Koji acid production by .4. wentii. On the whole, 
strains of the species appear to be somewhat less active biochemically than 
either the black Aspergilli or members of the A. flavus-oryzae group. It is, 
nevertheless, a vigorously growing species with definite biochemical pos- 
sibilities and should not be overlooked in any program relating to mold 
fermentation. 

Aspergillus alliaceus appears periodically upon alliaceous bulbs and 
occasionally upon cacti as at least a secondary parasite. It is not in- 
frequently isolated from soils, and appears to be fairly common in the 
southwestern states of Texas, New Mexico, and Arizona. It has also 
been isolated from soils of other areas including Southern Mexico and 
India. Nothing is known regarding its biochemical possibilities. 

Only the type strains of A. avenaceus and A. panamensis are known and 
neither has been shown to have any economic importance. 



Chapter XIX 
THE ASPERGILLUS TAMARII CxROUP 

Outstanding Characters 

Conidial heads radiate, generally loose-textured, hemispherical to globose, 
yellow-brown to olive-brown in color with green shades entirely lacking 
or only transiently produced in early stages. 

Conidiophores colorless, typically roughened throughout a part or all 
of their length. 

Vesicles subglobose to globose, fertile over the upper half to the entire 
surface. 

Sterigmata in one or two series depending upon the species and strain, 
often showing both conditions within the same head. 

Conidia heavy-walled, rough, elliptical, pyriform or subglobose, de- 
pending upon the species. 

Sclerotia commonly present, purple to reddish-purple or black, white- 
tipped when young. 

The Aspergillus tamarii group represents a collection of more or less 
closely related forms that are believed to be intermediate between the 
A. wentii and A. flavus-oryzae groups. Relationship to the latter group 
is unquestionable since there are intergrading forms which almost com- 
pletely bridge the gap from one group to the other. The group embraces 
two principal series: one, typified by A. terrieola, is characterized by dull 
yellow-brown conidial heads which never show any trace of green; the 
other, typified by A. tamarii, possesses dark brown conidial heads which 
commonly show transient shades of olive-green during the period of rapid 
growth. 

Group Key 
I. Conidia strongly elliptical, lemon-shaped A. citrisporus von Hohnel 

II. Conidia not strongly elliptical. 

A. Conidial heads light yellow-brown when mature, showing no green color at 
any stage. 

1. Colonies predominantly floccose, conidia with prominent projecting 

tubercles A. lulcscens Bainier 

2. Colonies not predominantly floccose. 

a. Heads large, conidia coarsely roughened with flattened bars and 

tubercles A. terrieola Marchal 

b. Heads small, conidia finely roughed 

A. terrieola var. americana Marchal 

250 



THE ASPERGILLUS TAMARII GROUP 251 

B. Conidial heads dull dark brown when mature. 

1. Green shades commonly evident, but confined to early stages 

A. tamarii Kita 

2. Green color persisting for several days, but eventually disappearing 

Bronze series 

Aspergillus citrisporus von Hohnel, in Sitzungsber. K. Akad. Wiss. Wien, 

Math. -Naturw. Kl. Ill, I Abt., p. 987, 1902. See The 

Aspergilli, Thorn and Church, pp. 191-2. 1926. 

Colonies on Czapek's solution agar, at room temperature, spreading 
fairly rapidly as submerged mycelium, producing a sparse aerial growth 
of conidiophores only; conidial areas at first yellow then gold and finally 
orange-brown (fulvus of Saccardo's Chromotaxia, approximately Mikado 
brown of Ridgway); reverse colorless. Conidial heads up to 500m in 
diameter, globose, radiate. Conidiophores 1 to 2 mm. long by 20 to 25m 
in diameter, with walls thin, about 1m in thickness, turgid and studded 
with granules when examined dry, finely roughened, often collapsing in age. 
Vesicles 30 to 50m in diameter, nearly globose, fertile over the whole surface. 
Sterigmata in one series, 8 to 12m by 3 to 4m, producing loosely radiating 
chains of conidia, at first yellow or golden then brown. Conidia lemon- 
shaped, 5 to 9m by 5 to 6m, rough from irregularly branching ridges of 
coloring substance between the inner and outer walls. Sclerotia or per- 
ithecia verbally reported by Thaxter but not seen by us. 

Diagnosis based on Thaxter's isolate (Thorn Xo. 4181.10). Thaxter 
gave no description. He obtained it several times from caterpillar dung. 
It grows and fruits more abundantly on Sabouraud's agar but it is difficult 
to keep viable in stock cultures. Additional collections include strains 
from caterpillar dung in Ann Arbor, Michigan, and Hanover, New Hamp- 
shire. 

Aspergillus lutescens Bainier nomen nudum; described by Thorn and 
Church, in The Aspergilli, p. 193. 1926. 

Colonies upon Czapek's solution agar rapidly growing and broadly 
spreading, floccose-woolly, at first white, becoming rusty-yellow as conidial 
formation begins and develops unevenly over the surface (fig. 68 A), 
finally becoming chestnut-brown when conidial areas are mature; reverse 
of colony pale yellow. Conidial heads radiate, hemispherical to subglobose, 
approximately buckthorn brown to Dresden brown (Ridgway, PI. XV), 
comparatively small, ranging from 100 to 300m in diameter. Conidiophores 
12 to 15m in diameter, varying greatly in length, mostly short, arising from 
the substratum, or as branches of aerial hyphae with walls pale yellowish 
and with pitting present but not conspicuous, not giving a rough appearance. 
Vesicles globose to subglobose (fig. 68 B), 20 to 40m in diameter. Sterigmata 






* 4k 
ma 







A 



D 







^S^]ri;^»^ 



0-' 



B o 











** 



E 



A 



c 



F 



Fig. 68. A-C, Aspergillus lutescens NRRL No. 426: A, Colonies on Czapek's 
solution agar showing extreme floccose habit and light sporulation, 10 days; B 
Single conidial head, X 500; C, Conidia, X 1200. D-F, Aspergillus temcola NRRL 
424: D, Single colony on Czapek's solution agar showing somewhat floccose but 
heavily sporing colony, 10 days; E, Conidial heads showing sterigmata in a single 
series, X 500; F, Conidia, X 1200. 

252 



THE ASPERGILLUS TAMARII GROUP 253 

in one series in smaller and crowded heads, up to 15 to 20/x by 4.0 to 5.0m; 
sterigmata often in two series in larger heads with primaries about 15 to 
18m by 4 to 5m, secondaries 12 to 14m by 4 to 5m. Conidia subglobose, 
varying from 5 by 7m to 8 by 9m, conspicuously roughened with prominent 
tubercles of color (fig 68 C). 

The species is known only in the type culture from the Bainier collection, 
NRRL Xo. 425 (Thorn Xo. 4040.478), and as a second strain, XRRL Xo. 
420, isolated in the Soil Microbiology Laboratory, Bureau of Plant Industry, 
Washington, D. C, about 1939. 

Aspergillus terricola Marchal, in Rev. Mycologique 15, Xo. 59: 101-103. 

1893. 

Colonies umbrinus; mycelial hyphae 3 to 5m in diameter, without anasto- 
moses; conidiophores hyaline, continuous or septate in age, 000 to 1,000m 
by 7 to 10m (whole depth of colony growth); vesicles subglobose, hyaline, 
39 to 50m, radiately covered with sterigmata; sterigmata in one series, 12 
to 15m by 4 to 7m; conidia umber (Sacc), ovate or elliptical then globose, 
rough, with colorless connectives. 

Description, from Marchal, of a culture isolated from soil in Belgium; 
not reported elsewhere, see variety below. 

Aspergillus terricola var. americana Marchal, cultural description by Thorn 
and Church, in Am. Jour. Bot. 8: 125. 1921. 

Colonies on Czapek's solution agar growing rapidly at room temperature, 
often somewhat floccose in central colony areas (fig. 68 D), ranging from 
shades near yellow ochre (Ridgway, PI. XV) when young, to Dresden brown 
or mummy brown in age, near Saccardo's umbrinus (PI. VII A); aerial 
growth largely consisting of crowded conidiophores; reverse uncolored. 
Conidial heads radiate, hemispherical to subglobose, loose in texture, 
consisting of comparatively few divergent chains of conidia, up to 200m in 
diameter. Conidiophores 300 to 000m in length by to 8m in diameter, 
with walls pitted. Vesicles globose to subglobose (fig. 08 E), up to 25m 
in diameter, fertile over the upper two-thirds or three-fourths. Sterigmata 
in one series, 7 to 10m by 2 to 4m- Conidia tuberculate (fig. 08 F) from the 
presence of color bars variously distributed between the outer and inner 
wall, ovate to nearly globose, from 3 by 5m up to 5 by 7m, usually about 
5.5m, occasionally 5 to 8m in diameter. 

Type culture XRRL Xo. 424 (Thorn Xo. 4838) isolated by F. M. Scales 
from redland soil in Georgia and discussed by Scales, in Jour. Biol. Chem. 
19: 459-472, 1914, under the name A. terricola Marchal. Scales' culture 
was submitted to Marchal, who designated the form as A. terricola var. 
americana Marchal, distinguished as follows: "The dimensions of the 



254 



A MANUAL OF THE ASPERGILLI 



vesicles 14 to 20m instead of 30 to 50m; of the sterigmata 5.6 to 10.5m by 
2.2m instead of 12 to 1 5m by 4 to 7m ; the spores only very delicately verrucose, 
separate your fungus from A. terricola." 




Fig. 69. Aspergillus tamarii. A, Colony growing on Czapek's solution agar 
characterized by heavy conidial production, strain XRRL No. 427, 10 days. B, 
Conidial heads, X 160. C, Single head, X 300. D, Single head further enlarged 
showing rough walls of conidiophore, and sterigmata in two series, X 500. 



Aspergillus tamarii Kita, in Centralb. f. Bakt. etc. 2 Abt., 37, No. 17/21, 
pp. 433-452. 1913. .Characterization of .4. tamarii Kita given by 
Thorn and Church in Am. Jour. Bot. 8: 118. 1921. See also Thorn 
and Church, The Aspergilli, p. 194. 1926. 

Colonies on Czapek's solution agar spreading broadly at room tempera- 
ture (fig. 69 A), with vegetative hyphae mostly submerged, fruiting areas 



THE ASPERGILLUS TAMARII GROUP 255 

at first colorless, then passing through orange-yellow shades to brown in 
old colonies, variously Isabella color, light brownish-olive, buffy-citrine, 
medal bronze, or raw umber (PI. VII B) (Ridgway, Pis. XXX, XVI, and 
IV, Column 19, and PI. Ill, Column 17); not showing true green, but often 
presenting a suggestion of green that is transient and limited to areas of 
young heads; reverse uncolored or occasionally pinkish. Conidial heads 
varying greatly in size in the same fruiting area, from more or less columnar 
to nearly, but not completely, globose and up to 300m in diameter, with 
radiating chains and columns of conidia. Conidiophores arising from 
submerged hyphae, up to 1 to 2 mm. in length, colorless, with walls be- 
coming abruptly thinner at the base of the vesicle, frequently showing 
irregular thickenings within, as a rule markedly rough or pitted throughout 
part or all of their length (fig. 69 D), sometimes appearing smooth or nearly 
so when examined in liquid mounts, but consistently rough or pitted when 
examined dry. Vesicles globose to subglobose, 25 to 50m in diameter 
(fig. 69 C), with fairly thin walls which frequently crush in mounts, fertile 
over almost the entire surface. Sterigmata, in one series in small heads, 
in two series in large heads; primary sterigmata commonly 7 to 10m by 3 to 
4m, becoming 20 to 35m long in gigantic heads, secondary sterigmata 7 to 
10m by 3m- Conidia ranging from more or less pyriform, through sub- 
globose to globose, conspicuously roughened from prominent tubercles and 
bars of orange-3 r ellow coloring matter deposited between the loose outer 
wall and the firm inner wall, commonly ranging from 5.0 to 6.5m in diameter, 
occasionally up to 8m- Sclerotia produced by many strains, usually purplish 
or reddish-purple, globose to pjniform with apex white. 

Species characterization is based upon Thorn and Church's culture No. 
4235.12 (NRRL No. 429) which was submitted to and identified by Kita 
as A. tamarii (see Thorn and Church, Am. Jour. Bot. 8 : 118. 1921). 

The organism described by Kita proved to be one of a great series repre- 
sented in our collection. It is common among cultures examined from 
North and South America, from Japan, China, India, and from Europe. 
The species has been found to be quite common in soil collected from 
many areas in the United States. 

The outstanding characters are orange-yellow to brown colonies; coarse, 
colorless conidiophores, usually roughened but with this character sometimes 
obscure; large, radiate, loose-textured, conidial heads; sterigmata in one or 
two series, commonly with single and double sterigmata in the same head; 
conidia more or less pyriform 5 to 8m in long axis with tubercles or bars of 
orange-yellow coloring matter between the inner and the outer cell walls. 

In preparing the manuscript for "The Aspergilli" (1926), Thorn and 
Church introduced into their general key, without name, on page 248 
under No. 279, a series of forms with morphology and general .appearance 



256 A MANUAL OF THE ASPERGILLI 

bridging the gap between .4 . tamarii Kita and the A . flams group. Without 
publication they referred to these as "The Bronze Series." These strains 
have the yellow-green color of A. flavus during the early stages of their 
development, but subsequently develop the yellow to brown colors of A. 
tamarii. Recognition of this border group is necessary since some strains 
of A. tamarii do not assume a definitely green color at any state in their 
development, while others show green as a transient character. Strains 
of .4. flavus, on the other hand, are typically characterized by the green 
to yellow-green colors. Furthermore, the conidia of A . tamarii are typically 
quite roughened, showing prominent tubercles or bars of coloring matter 
deposited between the outer and inner walls. In contrast, the conidia of 
A. flavus are less coarsely roughened and show more numerous and smaller 
tubercles or echinulations, as well as a greater tendency for the coloring 
substance to be generally diffused throughout the spore envelope. The 
forms under consideration show, in some degree, the coloration and spore 
characters of both groups and are believed to be truly intermediate be- 
tween A. tamarii and A. flavus. 

The fact that we received from Baarn in 1933, as Blochwitz's A. luteo- 
virescens Bloch. (Ann. Mycol. 31: 73-83. 1933) a culture (Thorn No. 
5345) which represented satisfactorily this intermediate series is not 
accepted as justifying the assignment of this name to the series, since the 
morphological characters displayed by the strain were so completely at 
variance with the original description, and since Blochwitz considered his 
species to be close to A. ustus. We question whether any sharp line of 
separation can be drawn between the two series because of the repeated 
appearance of intermediate forms. While we do not feel justified in 
assigning to these forms any specific designation, we do feel obligated to 
continue to call attention to their existence. We have at times considered 
the desirability of moving the whole A. tamarii complex over into the A. 
flavus-oryzae group, but this course has been abandoned since it was felt 
that to do so would introduce into an otherwise perfectly integrated 
group, a series of organisms whose relationship to them, while strongly 
suggested, is not proved, and which in its typical form would introduce 
discordant features. 

The species listed below are believed to represent probable synonyms: 

Biourge attached the manuscript name A. vulpinus to a member of the A. tamarii 
series (Thorn No. 4733.146) and contributed it to our collection, but it does not seem 
sufficiently different from the species to warrant separation. 

One strain of A. tamarii was found in the Bainier collection (Thorn No. 4640.397) 
as A. cacao, nomen nudum; another under the same name came from Pribram. 

A. gigas Spegazzini, Myc. Argent. V, in An. Mus. Nac. Buenos Aires Ser. B. Tome 
13: 424. 1911, was described from decaying coffee leaves in terms that suggest its 
relationship to A. tamarii. 



THE ASPERGILLUS TAMARII GROUP 257 

A. spadix Amons, in Archief voor de Suikerindustrie in Nederlandsch-Indie Jaarg. 
29, Deel 1, pp. 12-14. Jan.-June 1921. From the description this is a synonym of 
A. tamarii Kita. Colonies described as yellow-brown to deep brown, growing well 
one ommon laboratory media, without aerial mycelium; in reverse colorless; rice- 
colored to light violet at first, then light fuscous brown; conidiophores up to 2 to 3 
mm. by 8 to 9/x with walls about 0.9m thick and pitted or rough; vesicles globose, up 
to 50m in diameter, or almost clavate in small heads; conidia 5.5 to 7.2m, rough. 

Culture: Amons. Not studied by us. 

A. erythrocephalus B. and C, in Jour. Linn. Soc. (London), Bot. 10: 362. 1869. 
(See Fungi Cubensis Wrightiana, 1868; Type No. 642 in Curtis Herbarium de- 
posited in the Cryptogamic Herbarium of Harvard University, bears Wright's No. 
764. Part of the original material was removed by Dr. Farlow and given to Thorn 
for study.) 

Microscopic examination of this type specimen gives measurements as follows: 
conidiophores 45 to 70m in diameter, up to 2 mm. in length, with walls very heavy 5 to 
12m thick, varying from 5 to 6m in the broader part to 10 to 12m at the narrower base, 
pitted or roughened; vesicles up to 100m in diameter, nearly globose, fertile all over; 
head washed free from spores about 150m in diameter; sterigmata in two series, pri- 
mary 8 to 10m in length, secondary 8 to 9m in length ; conidia commonly 8 by 6m, ranging 
up to 8 to 12m by 5 to 9m, finely pitted or roughened with rather thin walls. Colors in 
the material are questionable on account of the age of the collection. 

Cultures: None. Type material only known. Placed between the A. tamarii and 
A.flavus groups. The amended description is offered due to the existence of a type 
specimen with very conspicuous characters under a name only very briefly described 
in 1869. When grown upon natural substrata such as grains, et cetera, conidiophores 
and heads of A. tamarii become very much larger than those ordinarily produced in 
culture media. This might account for this specimen which bears the name A. 
erythrocephalus B. and C. 

Occurrence and Economic Importance 

Of the species included in this group of brown-spored Aspergilli, only 
A. tamarii is in any sense widely distributed or common in nature. Asper- 
gillus lutescens is known only as the type culture and as a second isolation 
made in Washington, D. C, many years later. Aspergillus terricola has 
not been positively identified since its description, although the form with 
smaller heads and less coarsely roughened spores designated A. terricola 
var. americana by Marchal is occasionally encountered. Aspergillus 
tamarii is, however, a cosmopolitan mold upon vegetable material under- 
going slow decomposition and can be isolated from almost all soils examined. 
Like A. niger and A.flavus, it is more frequently recovered from warm and 
semi-tropical soils than from cool, temperate soils, although it occurs in 
the latter. The species commonly appears with A. flavus and A. oryzae 
as a constituent part of the "koji" used in the fermentation industries of 
the Orient. Certain strains apparently produce appreciable amounts of 
diastatic and proteolytic enzyme, while other strains are known to produce 



258 A MANUAL OF THE ASPERGILLI 

kojic acid. Gould reported this in 1938, and it was subsequently confirmed 
by A. J. Moyer (unpublished notes) for a strain isolated from Panama 
soil at the Northern Regional Research Laboratory in 1941. 

Kita's culture was isolated from a soybean sauce termed "Tamari", 
hence the species name. Tamari is made by a shorter fermentation process 
than soy sauce or shoyu, and differs from it in flavor. Ivita believed that 
where it was made empirically, it owed its individuality to the particular 
aspergillus which he isolated and described. 



Chapter XX 
THE ASPERGILLUS FLAVUS-ORYZAE GROUP 

Outstanding Characters 

Colonies varying from very light greenish-yellow to deep yellow-green 

(Ivy Green). 
Conidiophores rough or pitted, colorless. 
Heads hemispherical to columnar to subglobosc. 
Sterigmata in one or two series, often varying in the same head. 
Vesicles variable in form, from hemispherical to dome-shaped in small 

heads to globose in large heads. 
Conidia more or less roughened, varying in color as the colony. 
Sclerotia characteristic of many strains, generally grayish-brown to 

black, entirely lacking in others. 

Two species names are widely used for members of this cosmopolitan 
group. Aspergillus oryzae is applied quite generally, without regard to 
morphology, to the strains used by the Japanese and Chinese in the fermen- 
tation of rice and soy products. Although purified cultures are used in 
many places, the nomenclature is based more upon utilization than upon 
morphology. There appears, however, in these industries, a series of strains 
with long conidiophores, radiate heads, mostly greenish-yellow, with the 
green often fading completely in old cultures. These strains appear to be 
most commonly used in the production of the diastatic type of ferments and 
to be distributed in the great culture collections as Aspergillus oryzae 
(Ahlb.) Cohn. Such strains seem to be mostly oriental or tropical in origin. 
Strains with shorter conidiophores and yellowish-green heads, on the other 
hand, appear wherever fermenting or decaying materials are examined 
microscopically, or by culture. Aspergillus flavus Link has been accepted 
as a species aggregate for this second array of forms from which the segrega- 
tion of sections for description as separate species has been found difficult, 
if not almost impossible. If one wishes to perpetuate species names as 
roughly covering aggregates of closely related but varying strains, bearing 
always in mind that no sharp lines of differentiation exist, certain applica- 
tions of names may be made arbitrarily about as follows: 

Group Key 

I. Sterigmata mostly in one series, double sterigmata also present. 

A. Conidiophores long, 1-several mm., heads radiate, greenish-yellow; conidia 
pyriform, more or less roughened, variable in size up to 6, 8, or even 10ju 
in long axis A . oryzae ( Ahlburg) Cohn 

259 



260 A MANUAL OF THE ASPERGILLI 

B. Conidiophores 600 to 1700m; heads radiate, hemispherical, pale greenish- 

yellow; conidia smooth, globose 3.0 to 4.6^ A. micro -virido-citrinus 

Costantin and Lucet 

C. Conidiophores mostly less than 500/x; heads deep yellowish-green (Ivy 

Green); described as a parasite of the mealy bug of cane, occasionally 
elsewhere A . parasiticus Speare 

II. Sterigmata mostly in two series but single series common and often in same head, 
small heads usually showing single series only. 

A. Conidiophores very variable in length, mostly 400 to 1000m; heads in various 

yellowish-green shades A. flavus Link 

B. Conidiophores mostly borne as short branches from trailing hyphae forming 

an uneven cottony mass; heads white to yellow with traces of green only 

A. effusus Tiraboschi 

Literally hundreds of strains of this group have been collected and 
compared. Many of them have been isolated from fermentation investiga- 
tions in the laboratory and from industrial processes. No correlation of 
colony appearance, conidiophore or head morphology, color or microscopic 
detail, with actual utilization has been proved. A culture labeled A. 
oryzae (Ahlburg) Cohn, NRRL Xo. 447 (Thorn No. 113) has been preserved 
for over 30 years without apparent change in morphology. It is probably 
derived from Cohn's organism. When, however, we scrutinize the Asper- 
gilli obtained from the rice or soy fermentations of the Orient, cultures of 
the type represented by this strain are not the most common. The pre- 
eminently useful strains usually have the aspect of forms intermediate 
between A. flavus and A . oryzae. The dwarf green A . parasiticus of Speare 
isolated from dead mealy bugs of sugar cane in Hawaii proved no more 
parasitic to the same species of insects in the Barbados than other .4 . flavus 
strains sent with it. Teizo Takahashi contributed his series of strains under 
the letters used in his publication (1913). These are discussed at some 
length in Thorn and Church's paper on A. flavus, A. oryzae and associated 
species (1921), and also in "The Aspergilli" (192G, p. 202). It is sufficient 
to say that they vary all the way from almost white with few lightly colored 
heads to rich yellow-green in which heads are very numerous and fairly dark. 
They vary likewise in the length and diameter of their conidiophores. 
Characters of color and conidiophore length are not always correlated, 
although it is generally true that the darker conidial masses are borne upon 
shorter stalks. The collections contributed by Oshima, Kita, Hanzawa, 
and others from Japan as well as those isolated from commercial "Koji'' 
(sold as inoculum for fermentation industries) showed mainly the .4 . flavus 
morphology. Strains of this series appear constantly where cultures are 
made from soil or from decaying vegetation. A . flavus and its allies appear 
in collections from every correspondent who contributes Aspergilli. It is 
debatable whether the worker will be benefited or confused by the introduc- 



THE ASPERGILLUS FLAVUS-ORYZAE GROUP 261 

tion of some of the species names applied to members of the group. It 
must not be forgotten that any variant from the dwarf and deep green A. 
parasiticus to the longest stalked and palest greenish-yellow .4. oryzac may 
be found if we look for it. 

Aspergillus oryzae (Ahlburg) Cohn, in Jahresb. Schles. Gesell. Vaterl. 
Cultur (1183) 61: 226 Breslau. 1884. 

Synonym: Eurotium oryzae Ahlb. The name E. oryzae with an incom- 
plete description for the sake organism was published by 
Korschelt, in Dingier 's Polytechnisches Jour. 230: 330. 
1878, as taken from a letter from "Herr Ahlburg." See also 
Thorn and Church, Amer. Jour. Bot. Bot. 8: 106. 1921, and 
The Aspergilli, p. 198. 1926. 
Colonies on Czapek's solution agar rapidly spreading with vegetative 
hyphae mostly submerged and forming a white to gray mycelial layer in the 
form of a tough felty mass (fig. 70 A); developing pale greenish-yellow 
shades with the production of ripening conidial areas, varying from lime 
green to mignonette green (Ridgway, PI. XXXI, column 25) with the green 
disappearing later and the general color shifting to yellowish-brown shades; 
mycelium and agar uncolored. Conidial heads predominantly large, abun- 
dant, globose, radiate, with chains of conidia separate rather than adhering 
(fig. 70 D), giving the pale yellow shades of the colonies. Conidiophores 2 
to several mm. long by up to 20 to 25m in diameter with walls rather thin, 
definitely pitted or rough (fig. 70 F), colorless. Vesicles globose to sub- 
globose, less often hemispherical, up to 50 or even 70m with walls 1 to 1.5m- 
Sterigmata commonly in one series up to 15 or 20m long by 3 to 5m; or in two 
series with primary sterigmata up to 12 by 5m, and secondary sterigmata 10 
to 12m by 3.5m (fig. 70 B). Conidia more or less pyriform (fig. 70 F), vary- 
ing greatly in size in the same culture and in different strains, 3 by 4m, 4 by 
5m, 5 by u> or up to 9m or 10m hi long axis occasionally, rather thin-walled, 
roughened, becoming coarsely and deeply roughened in some strains. 
Sclerotia dark, few and not forming clumps, produced sporadically under 
undefined conditions. 

Diagnosis based primarily on culture XRRL Xo. 447 (Thorn Xo. 113) 
received from the Centraalbureau at Baarn and believed to be derived from 
Cohn's original strain. 

While the above description is believed to conform closely to the original 
conception of .4. oryzae, strains possessing the essential morphology de- 
scribed but which are heavier sporing and somewhat darker in color are more 
commonly encountered. Culture XRRL Xo. 458, obtained from Dr. 
Oshima as strain AoOld and shown in PI. VII C, and Fig. 70 C-F, is repre- 
sentative of these forms. 










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Fig. 70. Aspergillus oryzae. A and B, Strain XRRL Xo. 447 (Thorn No. 113): 
A, Single colony on Czapek's solution agar, loose-textured, conidiophores long and 
limited in number, 10 days; B, Details of single head, vesicle thin-walled, sterigmata 
mostly in two series, X 480. C-F, Aspergillus oryzae XRRL Xo. 458: C, Single 
colony on Czapek's solution agar, comparatively heavy sporing, 10 days; D, Conidial 
heads of the same, X 18; E, Conidial heads further enlarged, X 270; F, Single head 
showing vesicle, sterigmata in a single series, and roughened conidiophore, X 775. 



262 



THE ASPERGILLUS FLAVUS-ORYZAE GROUP 263 

The production of perithecia by members of this series was reported by 
Bezssonoff (1919) without adequate description and by Zikes (1922) whose 
culture, as received from him, belonged in the A. glaucus group. No 
ascosporic form is verifiable for the group thus far. 

Aspergillus micro-virido-citrinus Costantin and Lucet, in Ann. Sci. Xat. 

Bot. (IX) 2: 158. 1905. 

The appearance of colonies and measurements of conidiophores, heads, 
and spores indicate a form intermediate between A. flavus and .4. oryzae 
except for its small conidia. The description is very nearly satisfied by 
Takahashi's culture "P" (XRRL Xo. 480). It was found to grow between 
15° and 45° C. and to be pathogenic to rabbits. Colonies were greenish- 
yellow to predominantly yellow but contained some definitely green admix- 
ture in contrast to A. oryzae, which often lacks green color entirely. Co- 
nidiophores 600 to 1700m in length, up to 21m in diameter near the vesicle, 
uncolored, "granular" (= pitted) above, smooth toward the base. Vesicles 
24 to 62m in diameter. Sterigmata varying in size and arrangement with 
the size of the heads examined. Conidia globose, smooth, 3 to 4.6m (3.1m 
as a minimum to occasional diameters of 5.5m). 

An occasional culture shows the morphological characters described by 
Costantin and Lucet. Xo actual identity has been proved. 

Aspergillus flavus Link, in Obs. p. 16. 1809; also in Sp. Plant. 6: 66. 
1824, cited as synonym of Monilia flava Persoon, Myc. 1, p. 30. 

Synonym: Eurotium Aspergillus flavus DeBary and Woronin, in Beitrage 
zur Morphologic und Physiologie der Pilze, III Reihe, p. 
380. 1870. Exsiccati by Brefeld preserved in Rabenhorst, 
Fungi Europaei Edit. Xov. ser. II, Xo. 2135; one packet 
in the collection of the Xew York Botanical Garden. 

Colonies on Czapek's solution agar spreading rapidly, with floccosity 
limited to scanty growth of sterile hyphae in older and dryer areas among 
crowded conidiophores; conidial areas range in color in various strains from 
sea-foam yellow through chartreuse yellow, citron green, lime green, to 
Kronberg's green (PI. VII D and fig. 72 A), or even to ivy green, (See Ridg- 
way, PI. XXXI, column 25), yellow-green colors are either persistent or, in 
old colonies, altered by the disappearance of the green factor leaving shades 
of yellow-brown; reverse yellowish at first, passing over into brown shades 
in age. Conidial heads vary from small with a few chains of conidia to 
large radiate (fig. 72 E) or columnar masses in the same culture and varying 
mixtures of different types and sizes of head. Conidiophores mostly arising 
from submerged hyphae, commonly 400 to 1000m l°ng by 5 to 15m in diame- 



264 



A MANUAL OF THE ASPERGILLI 



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Fig. 71. Conidial structures in the Aspergillus flarus-oryzae group. A, A. flavus, 
NRRL No. 482: A 1; typical, large, radiate to globose head showing sterigmata in two 
series; A 2 , small, loosely columnar head showing single series of sterigmata. B, A. 
effusus, NRRL No. 506: B u large radiate head showing double series of sterigmata; 
B 2 , smaller head showing sterigmata in a single series; B 3 , diminutive head borne 
upon one of a chain of foot cells. In this group single and double sterigmata often 
occur in the same head (not illustrated). 






Fig. 72. Aspergillus flavus. A, Typical strain, NRRL No. 1957, showing crowded 
conidial heads and occasional black sclerotia. B, Heavy sclerotium producing 
strain, NRRL No. 500. C, Strain used for the production of kojic acid, NRRL No. 
484 (This approaches the character of A. oryzae). D, Dr. White's strain of A. flavus 
(NRRL No. 501) showing characteristic thin growth and sparse sporulation. E, 
Typical conidial heads, X 18. F, Single head showing vesicle, sterigmata, conidia, 
and the roughened conidiophore, X 780. All cultures on Czapek's solution agar, 
10 days. 

265 



266 A MANUAL OF THE ASPERGILLI 

ter, with walls pitted, rough (fig. 72 F) almost spiny in appearance, broaden- 
ing upward and gradually enlarging into vesicles 10 to 30 or 40m in diameter, 
dome-like in the smaller heads, flask-shaped in larger heads (fig. 72 F). 
Sterigmata in a single series in many smaller heads (fig. 71 A 2 ), or both single 
and double series on the same vesicles in large heads (fig. 71 Ai), varying 
from single sterigmata only 10 to 15m by 3 to 5m, to primary sterigmata 7 to 
10m by 3 to 4m and a secondary series 7 to 10m by 2.5 to 3m- Conidia pyri- 
form to almost globose, nearly colorless to definitely yellowish-green, vary- 
ing from 3m, 3 by 4m, 4 by 5m, or even larger and marked variously with pits, 
echinulations, or irregularly winding color bars and ridges to give a rough- 
ened effect of varying intensity. Sclerotia, when found, at first white then 
brown, hard parenchymatous, and a few strains white tipped, produced by 
some strains regularly and abundantly (fig. 72 B), scantily by others under 
undefined conditions. Perithecia not found. 

Description originally based upon culture NRRL No. 482 (Thorn No. 108) 
from the Centraalbureau at Baarn, Holland, but supplemented by obser- 
vation of many hundreds of cultures from many substrata and all parts of 
the world. 

Unless segregation under a specific name is supported by adequate 
morphological and reproducible cultural data, there is no way to identify 
the organisms intended. Applying these criteria, no reasons are seen for 
the use of the following specific designations: 

A. wehmeri Constantin and Lucet, in Ann. Sci. Nat. Bot. (IX) 2: 162. 1905. 

A. variabilis Gasperini, in Atti. Soc. Toscana Nat. Sci. Pisa Mon. 8, fasc. 2: 326. 
1887. 

A. pseudo-flavus Saito, in Centralb. Bakt. etc., 2 Abt., 18, No. 1/2, p. 34, figs. 15-18. 
1907, or its synonym S. pseudc-flava (Saito) Sacc, in Syll. 22: 1260. 1913. 

A. siebenmanni Constantin and Lucet, in Ann. Sci. Nat. Bot. (IX) 2: 162. 1905, 
is a bibliographic species based upon an organism isolated from the human ear and 
diagnosed by Siebenmann (Zeitsch. f. Ohrenheilk 12: 1883) as A. flavus. The de- 
scribes regarded it as a separate species based only upon the description given by 
Siebenmann. 

A. gymnosardae Yukawa, in Jour. Col. Imp. Univ. Tokoyo 1: 362, PI. 18, figs. 1-7. 
1911. A member of the A. flavus-oryzae group with measurements intermediate 
between more typical representatives of the two species. 

A. Ihomii Graff, nomen nudum, a heavy sclerotium-producing strain distributed 
by Graff but never described. No diagnostic basis for the name was presented. 

A. pollinis Howard, in Am. Bee Jour. 36: 577-578. 1896, was discussed as an or- 
ganism causing "pickled brood and bee paralysis" (See also idem. 38: 530-531. 1898). 
Turesson (Svensk Bot. Tidskr. 11: 30. 1917) decided the mold was A. flavus. 

Aspergillus parasiticus Speare, in Hawaiian Sugar Planters' Exp. Sta., 

Path, and Physiol. Ser. Bui. 12, p. 38, pi. 3-4. 1912. See 

Thorn and Church, The Aspergilli, p. 203. 1926. 

Colonies on Czapek's solution agar with sucrose spreading rapidly, 
forming a surface growth of crowded conidiophores with very few sterile 



THE ASPERGILLUS FLAVUS-ORYZAE GROUP 267 

hyphae (fig. 73 A), in deeper yellow-green shades near ivy green (Ridgway, 
PI. XXXI); reverse uncolored or yellowish. Conidial heads radiate, 
abundantly produced and giving color to the colony. Conidiophores given 
by Speare as 300 to 700m long, commonly under 400ju, with walls colorless, 
prominently rough or pitted, enlarging from 3/x at the foot up to 10 to 12/x, 
and passing into vesicles up to 35/* in diameter (fig. 73 B). Sterigmata in 
one series, 7 to 9/x by 2.5 to 3/x, closely packed over the vesicular surface, 
yellow. Conidia pyriform to globose, very rough, 4 to 5/x, occasionally 6/x 
in long axis, green. Xo sclerotia or perithecia reported. 

Described as parasitic upon the mealy bug of sugar cane (Pseudococcus 
calceolariac Mask.) in Hawaii. Type culture XRRL Xo. 502 (Thorn No. 
3509) received from Speare. Cultures with the same morphology Avere 
isolated from infected mealy bugs from Demerara by Thom, and in Louisi- 
ana by Kopeloff . Johnston, working in Puerto Rico, considered that he had 
proved infectivity to be a strain function among organisms of the A. flavus 
series rather than associated with morphology. Blochwitz (Ann. Mycol. 
32(1/2) : 8G. 1934) has called another nearly allied form A. flavus vsuwiridis, 
but gives no adequate data for separation. Cultures with these characters 
are occasionally obtained from sources not known to be associated with 
disease of insects. Shih has likewise described from China as Aspergillus 
chungii (Lingnan Sci. Jour. 15(3): 378. 1933) a strain which apparently 
duplicates .4. parasiticus. 

Aspergillus effusus Tiraboschi, in Ann. di Bot. (Rome) 7: 16, fasc. 1. 

1908. See also Thom and Church, in Am. Jour. Bot. 8: 109-110. 

1921, and Thom and Church, in The Aspergilli, p. 208. 1926. 

Colonies on Czapek's solution agar rapidly and broadly spreading, floc- 
cose or piled cottony white (fig. 73 C), becoming dirty yellowish or, in re- 
stricted areas, pale greenish-yellow, then passing over into dull buff or tan 
shades as heads mature ; reverse and agar yellowish. Conidial heads usually 
more or less columnar, mostly small, a few of them fairly large, many of 
them upon short conidiophores (fig. 71 B 3 and 73 E), often less than 100/x 
long and 5 to 10/x in diameter, arising from the trailing floccose hyphae, 
quickly losing their yellow-green color. Conidiophores with walls pitted or 
roughened, sometimes bearing granules (produced by drying droplets of 
exuded fluid). Vesicles mostly under 20/x in diameter (fig. 71 B 2 ). Sterig- 
mata in one series in small heads, in either one or two series in large heads 
(fig. 71 BO, approximating the A. flavus type. Conidia pyriform to 
globose varying from 3 by 4/x to 5 by 7m- Xo sclerotia or perithecia re- 
ported. The species was described originally from rotten corn (Zea 
Maijs). 

Culture description as given centers around culture XRRL No. 506 
(Thom Xo. 130) isolated by Dr. B. F. Lutman, Burlington, Vermont. Other 





T*iJ& 




B 




• 









i ^| 






' .if-- 




0& 



A 



■ 






00 



V 

■ 






v. 
I 



Fig. 73. A and 5, Aspergillus parasiticus, NRRL No. 465: A, Colony on Czapeks' 
solution agar showing heavy production of short-stalked conidial structures, 10 
days; B, Single conidial head showing single series of sterigmata and roughened 
conidiophore, X 500. C-F , Aspergillus effusus, NRRL No. 506: C, Colony onCzapek's 
solution agar showing characterisitc floccose habit and light sporulation, 10 days; 

D, Conidial structures arising from aerial hyphae characteristic of species, X 165; 

E, Single short-stalked fruiting structure arising from aerial hyphae, X 300; F, 
Conidial head-Showing sterigmata mostly in two series, roughened surface of conidio- 
phore not apparent, X 500. 

268 




THE ASPERGILLUS FLAVUS-ORYZAE GROUP 269 

strains studied include isolations from cornmeal from Indiana and from 
mealy bugs in Puerto Rico. Superficially this species bears little relation 
to A. flavus. Detailed microscopic examination of heads and spores, 
however, reveals true and close relationships. Selective transfer from 
original colonies permitted changes in the predominance of the sterile 
areas over the fruiting areas without changing the general habit or nature 
of the colony. Blochwitz (Bot. Centralb. Beiheft Abt. Anat. Phys. 48: 
176-182. 1931) regarded A. effusus as a floccose type of A. flavus. This 
position can be supported, but the authors feel that the species should be 
maintained since it is strikingly different from A. flavus in its general colony 
appearance, and since it is repeatedly, although infrequently, isolated from 
nature. 

Whether Sterigmatocijslis lulea Bainier (Bui. Soc. Bot. France 27: 27. 1880) 
was one of these can only be guessed from culture NRRL No. 508 (Thorn No. 
4640.473) received from the Bainier collection under this name. The strain is close 
to A. effusus. Bainier did not claim identity with S. lutea van Tieghem (Bui. Soc. 
Bot. France 24: 103. 1877), which was entirely undescribed. 

Aspergillus jeanselmei Ota, in Ann. de Parasit. 1(2): 137-146. 1923, as received 
from Baarn in 1939 (NRRL No. 507: Thorn No. 5665) represented a member of the 
A. flavus series with close affinities to A. effusus. 

Occurrence 

Members of the A . flavus-oryzae group are among the most abundant of 
all the Aspergilli. They are world-wide in distribution and are omnivorous 
in the substrata upon which they are able to grow and develop. They have 
been isolated from the widest variety of sources including : the fermentation 
industries of the Orient, grains and cereal products from different parts of 
the United States, various types of forage, egg noodles, bread and other 
bakery products, leather goods, dried dates, cured meats, dairy products, 
nut meats, soy sauce, home-canned fruits and vegetables, textiles, paper 
pulps, insects, tannin inoculum, feces, sputum, the lung of a bird, and from 
the duodenum of man. They are very abundant in soil, and have been 
observed in almost all samples examined. They appear to be particularly 
common in the warm soils from tropical and sub-tropical areas. They vary 
greatly in cultural appearance and in the detailed measurements of their 
fruiting structures, and to a limited degree these differences can be corre- 
lated with the sources from which they are obtained. Soil isolates com- 
monly show conidial heads near yellow-green in color which are borne upon 
comparatively short conidiophores. Isolates from the rice and soy fermen- 
tations of the Orient often show conidial heads pale yellow-green in color 
that are borne upon long, thin-walled conidiophores. Exceptions to this 
very general statement are common. 



270 A MANUAL OF THE ASPERGILLI 

Kojic Acid 

The ability of members of the A. flavus-oryzae group to produce kojic 
acid has been recognized for more than three decades. It is only within the 
past fifteen years, however, that serious attention has been given to this 
fermentation. Beginning with the work of Challenger, Klein, and Walker 
in 1929 and 1931, and continuing with that of May, Herrick, Moyer, Ward, 
and Wells in 1931 and 1932, the proper nutrients and cultural conditions 
necessary for its production were defined. Subsequent contributions have 
been made by Kluyver and Perquin (1933) and by Barham and Smits 
(1936). In all of the early reports the responsible cultures were cited as A. 
oryzae, whereas in more recent ones the cultures employed have generally 
been identified as Aspergillus flavus. It is of interest to note that the strain 
studied by May and associates (NRRL No. 484: Thorn No. 3538) was a 
thoroughly typical A . flavus when first isolated by Thorn in 1914, but during 
the long period that it has been maintained in artificial culture it has gradu- 
ally changed until today it more nearly resembles A . oryzae in its general 
habit and coloration (fig. 72 C). Its capacity to produce kojic acid remains 
undiminished, however. The culture employed by Barham (NRRL No. 
625) likewise fails to satisfy the typical cultural picture of A . flavus, although 
it is discussed under this name. In contrast to these cultures, other strains 
belonging to this group have been under continuous laboratory cultivation 
for more than 30 years without apparent change in appearance or behavior. 
The above and additional references to the kojic acid fermentation are pre- 
sented in the Topical Bibliography, pp. 297-298. 

Enzymes 

Members of the A . flavus-oryzae group produce diastatic and proteolytic 
enzymes abundantly. For this reason they have been much studied, and 
an extensive literature regarding mold enzymes has developed around the 
use of these fungi. In large measure the alcoholic and soy food industries 
of the Far East are based upon these molds and their enzymes. In the 
production of alcoholic beverages, the diastatic enzymes produced by an 
Aspergillus (regularly identified as A. oryzae) are employed to hydrolyze 
the rice starch. Alcohol is then produced from the resultant sugars by the 
addition of a fermentative yeast. In the soy industries, closely related 
molds, or even the same strains, are used as a source of proteolytic enzymes. 
In 1894 Takamine secured a series of U. S. patents covering the production 
of diastatic enzymes and the making of alcoholic liquors (see Topical 
Bibliography, p. 302). Subsequent to this, other investigators, mostly 
Japanese, published a number of papers in this field. Oshima in 1922 and 
1928 reported on the production of protease by members of the A. flavus- 



THE ASPERGILLUS FLAVUS-ORYZAE GROUP 271 

oryzae group. Today considerable quantities of diastatic enzymes, proteo- 
lytic enzymes, and mixed diastatic and proteolytic preparations are being 
manufactured from these molds for use in the textile and tanning industries 
particularly. Within recent years considerable attention has been given 
to "moldy bran" (bran seeded with selected strains of A. oryzae) as a pos- 
sible substitute for malted barley as a saccharifying agent in the production 
of industrial alcohol '(Underkofler, Fulmer, and Schoene, 1939; Schoene, 
Fulmer, and Underkofler, 1940; Hao, 1942; Hao, Fulmer, and Underkofler, 
1943; Christensen, 1943). 

References to papers dealing with the production of enzymes by molds 
belonging to this group are presented in the Topical Bibliography, pp. 
302-304. No attempt has been made to present a complete bibliography 
of the subject, but it is believed that sufficient citations are listed to intro- 
duce the reader to the extensive literature of the field. 

Pathogenesis (See Topical Bibliography pp. 307-310) 

Pathogenesis has been occasionally reported for strains identified as mem- 
bers of the A. flavus series. Observations reporting their presence in the 
external ear go back to Siebenmann (1882). Ota described A. jeanselmei 
as a parasite of human nails in Paris in 1923. Bereston and Keil (1941) 
described a case of infected nails in which the strain, as seen by us, proved 
to be a variant of A . flavus. There are ample records to show an occasional 
infection of the human being. There are no data as to the route of infection, 
and the question whether the parasite is a primary or a secondary (wound) 
parasite stands unanswered. The constant presence of members of this 
group in every human environment, together with a lack of evidence of 
ability to penetrate sound human tissue, leaves some doubt — not as to its 
ability to grow when once established, but whether it can actually "break 
and enter" as a direct agent of disease. Aspergillus flavus is commonly 
isolated from sputum. In birds, cases of lung involvement have been re- 
ported. There is no question but that the mold can persist for reasonable 
periods of time within the animal body. A. flavus is one of the more com- 
mon air-borne molds, and occasional allergic reactions are attributed to it, 
although instances where it is the sole responsible agent are not known to 
have been reported. 

Antibiosis 

The production of antibacterial substances by strains of Aspergillus flavus 
has been observed by a number of workers during the past five years. 
White (1940) reported a culture of A. flavus (found by the writers to be a 
somewhat atypical strain) to produce some substance which was definitely 
bactericidal against some gram-negative and some gram-positive bacteria. 



272 A MANUAL OF THE ASPERGILLI 

A more detailed study of this strain was subsequently made by White and 
Hill (1943) and the name "aspergillic acid" was assigned to the active 
substance. The compound shows a comparatively high toxicity to labora- 
tory animals. Utilizing the White strain, Jones, Rake, and Hamre (1943) 
have made additional studies on the biological properties of aspergillic acid. 
In the meantime, Glister (1941), at Oxford University reported the produc- 
tion of an antibacterial substance effective against gram-negative and 
gram -positive bacteria by a different strain of A . flavus (found by the writers 
to be wholly typical of the species). He noted the possibility of relationship 
between the substance with which he was working and that earlier reported 
by White. 

Following the work of Jones, Rake, and Hamre (1943), McKee and Mac- 
Phillamy (1943) succeeded in demonstrating the production of a second and 
entirely different antibacterial substance. In certain chemical properties, 
and in its action on bacteria, this was found to resemble penicillin very 
closely, but actual identity was not proved. In a subsequent and more 
detailed report, McKee, Rake, and Houck (1944) defined more exactly its 
bactericidal action against various gram-negative and gram-positive bac- 
teria and proved additional evidence of its penicillin-like characters. They 
designated the substance i ''flavicidin'". 

Concurrent with this work, Bush and Goth (1943a and 1943b), working 
at Vanderbilt University, succeeded in demonstrating the production of an 
antibacterial substance from still another strain of A. flavus which was 
strongly active against Staphylococcus and other gram -positive forms but 
comparatively inactive against gram-negative forms belonging to the E. 
coli group. The substance was termed "flavicin". Identity with flavicidin 
and with penicillin is possible but has not yet been proved. Cook and 
Lacey (1944) report the production of appreciable amounts of an antibiotic 
substance from a strain of A. parasiticus. This was provisionally desig- 
nated "parasiticin", and its similarity to penicillin was noted. Identity 
with flavicin (Bush and Goth) and flavicidin (McKee, Rake, and Houck) 
was suggested. Since A. parasiticus is so closely related to A. flavus, it 
would seem probable that an antibiotic produced by it would be similar to 
that produced by the latter species under the same conditions. 

Waksman and Bugie (1943) investigated a large number of strains be- 
longing to the A. flavus-oryzae group. They found strains of A. oryzae to 
show little activity, whereas strains of A. flavus showed increased but vary- 
ing amounts. Yields' were markedly influenced by various nutritional and 
environmental factors. Two types Of antibacterial substances were ob- 
served: aspergillic acid and a substance similar to, if not identical with, 
penicillin. When grown in submerged culture, one strain was found to 
produce amounts comparable to the best strains of Penicillium notatum 
tested. Unfortunately, yields were not quantitatively determined. 



Chapter XXI 
THE ASPERGILLUS OCHRACEUS GROUP 

Outstanding Characters 

Conidial heads ranging from sulphur yellow to varying shades of ochra- 
ceous, depending upon the species and strain, showing a greenish tint 
only in the single species A. sparsus; heads globose or radiate with 
conidial chains commonly adhering into divergent columns. 

Conidiophores normally showing shades of yellow in the outer layers of 
the wall which is rough or pitted, usually prominently but occasionally 
reduced to traces which are seen most readily in dry mounts. 

Sterigmata in two series with the primary often quite large and septate. 

Conidia in some series thin -walled and smooth, in others showing defi- 
nitely double walls, more or less roughened or echinulate. 

Sclerotia present in most species and strains, often dominating the cul- 
tures; in others entirely lacking. When present, ranging in color from 
cream or buff through pink and orange shades to purplish-vinaceous. 

Molds belonging to this group are common wherever organic matter is 
decomposing under natural conditions. In spite of great variation in super- 
ficial appearance, length of conidiophore, size of heads, intensity and shades 
of color, and sclerotium production, they fit together into a great natural 
group of related forms. Extreme variants in the several series can be easily 
considered separate species and have been so described, but collections of 
great numbers of such forms present so many gradations that identification 
by description becomes doubtful if not impossible. Allocation to series 
centered upon some described species gives a practical method of grouping 
together closely related members of the great aggregate. 

The name "ochraceus", derived from the pigment ocher and attached to 
the most abundant series in the group, is an old mycological usage which was 
more or less definitely followed in Saccardo's use of it for a plaque in his 
Chromotaxia. Ridgway analyzed the colors more exactly, dropped the 
term ochraceus but included a plaque near to it as PI. XV, Col. 15 YO, 
Ochraceous-Buff . The strains of this group, however, mostly show colors 
closer to the yellower tints in Ridgway's Plate XXX, column 19. It is im- 
portant that relationship with the great group shall be quickly grasped 
whether exact identity with particular strains already known is claimed or 
not. 

Group characterization : The colony appearance of this group of Asper- 
gilli varies greatly with the presence or absence of sclerotia. Some species 

273 



274 A MANUAL OF THE ASPERGILLI 

are typically ochraceous in color from abundant conidial heads and are not 
known to produce sclerotia. Others produce a few sclerotia, or clusters of 
sclerotia, under very special conditions but still are characterized by their 
abundant conidial heads. Others always produce sclerotia in sufficient 
number to dominate the colony. Sclerotia, when present, are globose or 
elliptical, 0.5 mm. or more in long axis, and vary in color from shades of 
yellow to orange-vinaceous or even purplish. The submerged mycelium 
varies from colorless to yellow, orange, or purplish shades. Heads are 
globose or radiate and vary in color from bright yellow near citrine through 
pale yellows to varying shades of ochraceous but never appear in umber 
shades, and show green (near olivine, Ridgway, PI. XXXII) only in A. 
sparsus. Conidiophores vary greatly in length and diameter but have 
walls yellow, especially in the outer layer, pitted or rough — in occasional 
dwarf strains with fairly thin walls the pitting becomes difficult to demon- 
strate, hence careful examination with a good oil immersion objective may 
be necessary. Vesicles are globose, or occasionally somewhat elliptical, 
varying greatly in size with walls usually colorless and much thinner than 
the conidiophore wall. Sterigmata are always in two series, with primary 
sterigmata varying, as in the A. niger group, from fairly short to very long, 
but commonly 15 to 30m upon Czapek's solution agar and occasionally much 
longer on special substrata; secondary sterigmata differ less conspicuously 
than primary sterigmata, being commonly 7 to 10/x by 1.5 to 2.5m; both 
series of sterigmata are almost colorless when examined with high magnifi- 
cations. Conidia are small, sometimes elliptical, but mostly globose or 
subglobose, smooth in most strains, but in others very delicately wrinkled 
or spinulose without prominent tubercles or bars of color, mostly 2 to 4/x in 
long axis, larger in occasional strains. 

The following key is an attempt to arrange this group into a satisfactory 
order to bring nearly related organisms together. For this purpose specific 
names already in the literature with the particular distinguishing marks cited 
in their diagnoses are introduced as more or less definitely fixed points in 
the group with which new material or unknown material can be compared. 

Group Key 

I. Conidial heads in fairly pure yellow tints such as sulphureus and citrinus 

The A. sulphureus series 

A. Sclerotia usually not abundant; conidiophores up to 1000m long; heads 

abundant, radiate; vesicles globose; conidia globose or subglobose, 
smooth, about 3m A. sulphureus Fresenius 

B. Sclerotia abundant, commonly characterizing the culture; heads scattered, 

or grouped in restricted areas, particularly at the drying margin of agar 
slant cultures; conidia smooth, up to 4.0m 

A. quercinus (Bainier) Thorn and Church 
II. Conidial heads in pale to darker ochraceous shades. . . The A. ochraceus series 



THE ASPERGILLUS OCHRACEUS GROUP 275 

A. Conidia thin-walled, smooth A. ochraceus sub-series glaber 

1. Sclerotia dominating the colony appearance; conidia small, about 3.0m- 

a. Sclerotia abundant, buff to pink or purplish in color in age; heads 

hemispherical to columnar, cream-buff to pale ochraceous 

A . sclerotiorum Huber 

b. Sclerotia abundant, at first gray, quickly becoming black; heads 

globose or radiate; upon onions and other bulbs 

A. alliaceus Thorn and Church 
(A. uentii group, p. 241) 

2. Sclerotia present but secondary to the crowded, honey-yellow conidial 

heads; conidia smooth A. melleus Yukawa 

B. Conidia with firmer walls, echinulate or barred 

A. ochraceus sub-series echinulata 

1. Conidia elliptical to subglobose. 

a. Conidia 3.5 to 5.0m in long axis; echinulate heads ochraceous, 

radiate, and splitting; sclerotia in type material 

A. ochraceus Wilhelm 

b. Conidia 3.0 to 3.5m, spinulose; sclerotia not found 

A. elegans Gasperini 

c. Conidia up to 6.3m in diameter, rough (Otherwise A . ochraceus) 

S. buturacea Bainier 

2. Conidia globose, echinulate, 7.0 to 8.0m in diameter (Otherwise A. 

ochraceus) A. delacroixii (Sacc.) Thom and Church 

3. Conidia strongly elliptical or pyriform, spinulose, 4.0 to 6.0m by 3.0 to 

4.5 M A. ostianus Wehmer 

III. Conidial heads commonly showing a definite greenish tint, near olive-buff to 
olivine in color A. sparsus Raper and Thom 

Aspergillus sulphureus (Fres.) Thom and Church, in The Aspergilli, p. 185. 

1926. 

Syn.: S. sulphured Fresenius, in Beitr. zur Mykologie, Heft 3, p. 83, taf. 
XI, fig. 30-33. 1863. See also Sacc. Sylloge 4: 73. 1886. 

Colonies upon Czapek's solution agar at laboratory temperature, growing 
rapidly, forming a variously wrinkled to zonate felt, white or tinged with 
yellow to pink or purplish shades. Conidial heads irregularly produced in 
most cultures, in pale or sulphur yellor tints, mostly globose, but often split- 
ting into spore columns in age. Conidiophores with walls firm, rough or 
pitted, yellow, varying greatly with the strain from very short and sparingly 
produced to clustered in areas and dominating the culture, or entirely cover- 
ing the mycelium where sclerotia are absent. Vesicles typically globose and 
fertile over the whole surface. Radiating primary sterigmata closely 
packed on the vesicular surface, varying from 8 to 10m by 3 to 5/x to several 
times larger in some big heads, secondary sterigmata usually uniform, 
phialiform about 8 to 10m by 2 to 3m- Conidia small, globose or subglobose 
2 to 3 or 3.5m in long axis, thin-walled, smooth. 

A . sulphureus as described by Fresenius had long conidiophores and yel- 



276 A MANUAL OF THE ASPERGILLI 

low globose heads. Sclerotia were not reported. In our experience, how- 
ever, they are seen not infrequently in cultures which otherwise fit the 
description of A. sulphureus as presented. In the sub-series, represented 
by A. quercinus (Bainier, Thom and Church), sclerotia are very abundant 
and heads comparatively few and scattered among the sclerotia. In addi- 
tion to these two names which roughly designate sections or series of 
isolates as we find them, every gradation between these extremes may be 
anticipated. 

The species is fairly common in soils, on cereal grains, and upon decaying 
vegetation. 

Several species have been described as having conidial heads sulphur 
yellow but doubtfully separable from A. sulphureus. 

S. ochroleuca Spegazzini, in Myc. Argent V, p. 434, in Anal. Mus. Nac. Buenos 
Aires, Ser. 3, T. 13. 1911. 

S. auricoma Gueguen, in Bui. Soc. Myc. France 16: 171-187, figs. 1-48. 1899. This 
appears to have been a member of the series with primary sterigmata proliferating 
abundantly to form little conidiophores and secondary heads thus giving the head the 
appearance of bearing yellow hair. Such proliferation occasionally occurs in many 
species but has not again been found in this group. 

S. vitelline, Ridley, in Jour. Bot. (London) 34: 152, pi. 257, figs. 14-16. 1896, is 
described as producing bundles or coremia composed of partially adherent conidio- 
phores. The organism does not appear to have been cultivated and has not since 
been reported. 

Aspergillus quercinus (Bainier) Thom and Church, in The Aspergilli, p. 186, 

1926. 

Synonym: S. quercina Bainier, in Bull. Soc. Bot. France 28: 78. 1881. 
See also Sartory, Etude biologique du Sterigmatocystis 
quercina Bainier, in Bui. Soc. Myc. France 26: 349. 1910. 

Colonies upon Czapek's solution agar spreading broadly, characterized 
by the presence of an aerial white mycelium and the abundant production 
of sclerotia over the whole area (PI. VII, E and fig. 74 A), or in sectors, or 
variously distributed ; the mass changing from yellow to orange-yellow and 
finally to rufous or brick red shades with the ripening of the sclerotia; re- 
verse in shades of yellowish-orange. Conidial heads in yellow tints, near 
sulphureus, scattered among the sclerotia or occurring in long-stalked 
groups in the dryer areas of the culture tube or plate, mostly up to 200> 
in diameter, but occasionally larger, up to 300 or even 400/x (fig. 74 C). 
Conidiophores with walls mostly pale yellow, especially in the outer layer, 
pitted (fig. 74 D), occasionally with abundant granules, about 2/jl in thick- 
ness, varying from short and inconspicuous in crowded sclerotial areas to 
very long tufts in dryer areas, up to 2 or even several millimeters by 10 to 
20m- Vesicles colorless, crushing easily, 35 to 45^ in diameter, fertile over 








* <vC jlU 


^^!E§3i?^* 


l**v#**otw 




^RWl' 


D 











■ * -*r ^ 


•^§ 


"^ 




> 


r 




,4 


| ¥■$ •"' 


■ 


IHf 



E 



Fig. 74. A-Z), Aspergillus quercinus, NRRL No. 394: A, Colonies on Czapek's 
solution agar, 10 days, predominantly sclerotial but with localized areas of heavy 
conidial production in central areas. B, Marginal area of the same colony showing 
very abundant sclerotia, and scattered conidial heads in lower right-hand corner. 
C, Conidial heads on hay infusion agar plate, X 18. D, Single head showing globose 
character, crowded sterigmata, and roughened conidiophore. E, Aspergill 
tiorum on Czapek's solution agar, 10 days. F, Portion of the same enlarge( 
abundant large sclerotia and scattered comparatively small heads, X 6. 

277 



us sclero- 
rged to show 



278 A MANUAL OF THE ASPERGILLI 

the entire surface. Sterigmata in two series: primary 10 to 20 or even 30m 
in larger heads by 2 to 4 or 7 m at the tips; secondary 10 by 2 to 
2.5/x. Conidia 2.5 to 3m by 3 to 3.5/* to almost globose, very nearly colorless 
in mounts, smooth, thin-walled. Sclerotia white to brick red, up to 500m 
in diameter (fig. 74 B). 

The original culture as described by Bainier in 1880 was not preserved. 
Neither was that of Sartory in 1910. However, cultures of organisms vary- 
ing about as here described and complying fairly closely with Bainier's 
description are not infrequently encountered from widely separated regions. 

Aspergillus sachari Chaudhuri and Sachar, in Ann. Myc. 32: 95. 1934. 

This organism was placed by the describers near ^4. quercinus on the basis of the 
colors of the head and sclerotia. They ignored the colorless, smooth conidiophores 
which, if correctly observed, would make their placing untenable. It is left here 
arbitrarily until rediscovered and its characters verified. Compare: A. candidus 
group. 

Species characterization based upon the original description follows: 

Colonies on Czapek's solution agar naphthalene yellow, (Ridgway, PI. XVI. 23, 
yellow, f.) more or less floccose; reverse colorless at first then in yellow shades. Scle- 
rotia scattered through the colony, quickly developed, hard, white through yellow 
shades to cinnamon, up to 2 mm. in long axis with a depression in the center. Heads 
abundant, radiate, 50 to 85m in diameter, or somewhat columnar in age up to 160 by 
100m. Conidiophores 700 to 9C0m by about 8m with smooth colorless walls, about 1.5m 
thick; vesicles globose, 20 to 30m in diameter; sterigmata in two series, covering the 
whole vesicle, radiate; primary 5.4 by 2m, secondary 7.2 by 1.8m, conidia colorless, 
smooth, globose, 2 to 2.7m in diameter. Found in alkaline soil. 

Aspergillus sclerotiorum Huber, in Phytopathology 23 (3): 
306-8, Fig. I. 1933. 

Colonies on Czapek's solution agar rather slow growing, forming a smooth 
mycelial layer irregularly overgrown with loose tufts of sterile hyphae within 
which numerous sclerotia develop and often dominate the colony appear- 
ance (fig. 74 E), white to shades of yellowish or ochraceous; reverse cream. 
Conidial heads given as sulphur yellow by Huber, but in our cultures found 
in ochraceous shades such as cream-buff (Ridgway, PI. XXX), mostly 
appearing as columnar masses of conidia up to 250m long, sometimes 
splitting, 60 to 70m at the base and up to 125m at the apex. Conidiophores 
with walls yellow and pitted, commonly 200 to 400m long by 7 to 10m in 
diameter, but varying from very short, 50m in length, to occasional groups 
up to 1200m; in old cultures, arising as branches from loose aerial hyphae. 
Vesicles subglobose, 15 to 20m in long axis. Sterigmata in two series, pri- 
mary usually 8 to 9m, occasionally much more but not above 20m long by 3.5 
to 4.5m in diameter, secondary 8 to 9m by 2m- Conidia smooth, mostly 2 
to 2.5m- Sclerotia abundant (fig. 74 F), beginning to appear within 3 days, 
white to cream or pink, up to 1.5 mm. in diameter, well scattered over the 



THE ASPERGILLUS OCHRACEUS GROUP 279 

mycelium or vaguely in zones, giving the characteristic appearance to the 
colony, hence the name. 

The type culture NRRL Xo. 415 (Thorn No. 5351 received from Huber) 
was isolated from rotting apples in Oregon and proved capable of causing 
decay in apples under controlled conditions in which other strains failed to 
produce rot. It has not been discussed by others. Blochwitz (Ann. 
Mycol. 32(1/2) : 88. 1934) considered A. sclerotiorum to be a synonym for 
A. clegans Gasperini, but this is not consistent with the description. 

Aspergillus melleus Yukawa, in Jour. Coll. Agr. Imp. Univ. 
Tokyo 1, No. 3, p. 366, Taf. 16. 1911. 

Colonies on Czapek's solution agar rapidly growing and spreading, form- 
ing an aerial felt of sterile hyphae and conidiophores, and producing 
sclerotia with walls white to yellowish-brown, commonly a shade of yellow- 
orange near "melleus" of Saccardo's Chromotaxia (Approximately Ridgway 
PI. XXX, 19", honey yellow; or warm buff, PI. XV, 17'd); reverse reddish 
to brown shades. Conidial heads ranging from small to large and globose, 
commonly splitting into dense columnar masses, sometimes 250 to 300m 
in long axis. Conidiophores with walls yellow, pitted, up to 500 or even 
1000/x long, 15 to 20 or even 25m in diameter. Vesicles up to 40 to 50m in 
diameter, globose in large heads, more or less pyriform in small heads, 
fragile and readily crushed in mounting. Sterigmata in two series, primary 
10 to 20/x by 2.5 to 4/x, secondary commonly 3 to 10m by 2 to 3/x, with oc- 
casional larger sterigmata in either series. Conidia in long chains, almost 
colorless when mounted, very thin-walled, smooth, slightly elliptical, about 
3m or a little more in long axis. Sclerotia ranging from 400 to 700m in 
diameter, in yellow-brown shades. 

Hanzawa furnished Thorn's culture No. 4291.6 (NRRL No. 416) as the 
type strain used by Yukawa. Other strains, including cultures from For- 
mosa, South Africa, and the United States, indicate that the cultural aspect 
of Yukawa's species is carried by organisms widely distributed. Some re- 
lated strain may have been described by Tiraboschi as A. ochraceus var. 
microspora (in Ann. di Botanica VII. 14. 1908) from corn bread in Italy. 
This name is used also by Nakazawa (Jour. Agr. Chem. Soc. Japan 10: 
English summary p. 185. 1934). 

Aspergillus ochraceus Wilhelm, in Inaug. Diss. Strassburg, p. 66. 1877. 
Wilhelm's exsiccati exist as Rabh. Fl. Europaei no. 2361. 

Colonies upon Czapek's solution agar spreading fairly broadly, usually 
plane, zonate in some strains, characterized by a tough submerged felt which 
may be colorless, yellow, orange, or purplish and, arising from this, more or 
less crowded conidial structures which give to the colony its characteristic 



280 



A MANUAL OF THE ASPERGILLI 



color and appearance (PI. VII F and fig. 75 A) ; reverse ranges from colorless 
through orange to purplish shades. 

Conidial heads when large mostly globose or variously splitting into 
masses of conidial chains (fig. 75 C), variously colored from very pale to 
deep ochraceous shades . Conidiophores vary greatly in length and diameter 




Fig. 75. Aspergillus ochraceus series. A and B, A. ochraceus, NRRL No. 398 
and No. 408, respectively, growing on Czapek's solution agar at room temperature, 
10 days. C, Mature heads of strain No. 398; the tendency to split into divergent 
columns in age is characteristic. D, Photomicrograph of a single head showing globose 
vesicle, sterigmata in two series, and coarsely roughened conidiophore, X 325. 

in the various strains, but typically show a yellow color in the outer layers 
of the thick wall which shows characteristic pitting or roughening (fig. 
75 D). Vesicles mostly globose or somewhat elliptical, and fertile over the 
whole surface (fig. 75 D). Sterigmata in two series: primary varying from 
small to very large, commonly 15 to 30/x in length; secondary fairly uniform, 



THE ASPERGILLUS OCHRACEUS GROUP 281 

commonly 7 to 10/x by 1.5 to 2.5/x. Conidia globose, subglobose, or some- 
what elliptical, more or less rough or echinulate, ranging from 3.5 to 5.0m 
in long axis. No perithecia reported. Sclerotia are present in Wilhelm's 
material preserved in exsiccati. 

Among the great number of isolates belonging to this group a considerable 
number show approximately the characters as drawn from Wilhelm's 
description and his exsiccati. In the limited sense then, ^4. ochraceus 
Wilhelm can be interpreted to apply to those ochraceous strains which bear 
echinulate conidia from 3.5 to 5.0m in diameter and produce sclerotia. 
Among the strains included, almost all are found to produce colonies of 
consistent aspect in continuous culture. 

Aspergillus elegans Gasperini, in Atti. Soc. Toscana Sci. Nat. Pisa 

Mem., 8: 328, fasc. 2. 1887. 

Synonym: S. elegans (Gasp.) Sacc, in Syll. 10: 525. 

This species by description differs little from A. ochraceus Wilhelm 
except for the absence of sclerotia, and conidia which do not exceed 3.5/x 
in diameter. Thorn and Church did not recognize it as a valid species in 
1926, and one may be certain that no sharp line can be drawn separating 
A. elegans from A. ochraceus. Nevertheless, forms producing conidia con- 
sistently less than 3. 5m are commonly encountered among miscellaneous 
isolations from nature, and there is an argument for retaining a species to 
include such forms. The following species diagnosis, taken from Saccardo 
(10: 525) was presented by Thorn and Church (1926): 

"Mycelium white; stalks continuous, unbranched, hyaline then pale 
ochraceous, 1 to 6 mm. long, by 5 to 12/x in diameter, delicately studded with 
drops ; vesicle up to 70m diameter, radiate, entirely covered with sterigmata; 
sterigmata, primary 4 to 26m long, secondary 7 to 14m long by 1 to 2m; 
conidia ochraceous, elliptical to globose, up to 3 to 3.5m, with wall very deli- 
cately verruculose, sclerotia not found." 

Various other species have been described which are obviously closely 
related and belong to the A. ochraceus series. A partial list would in- 
clude : 

S. helva Bainier, in Bull. Soc. Bot. France 28: 78. 1881. Thorn's culture No. 
4640.476, received under this name, came from the Bainier collection. 

A. alutaceus Berkeley and Curtis (in Grevillea 3, No. 25, p. 108. 1875) was the 
name proposed for a mold found upon corn. The specimen is preserved as No. 3793 
in Curtis' Herbarium now in the Cryptogamic Herbarium of Harvard University. 
The specimen shows that this species was probably a strain of A. ochraceus. 

A. ochraceus var. microspora Tiraboschi (in Ann. di Bot. [Rome] 7: 14. 1908) 
represents a strain in which all measurements were reported as reduced. 

A. rehmii Zukal. A culture from Dr. Westerdijk, received under this name, is also 
a member of this series. 



282 A MANUAL OF THE ASPERGILLI 

Certain large-spored species of doubtful validity and relationship have 
been described that are believed to represent members of the A. ochraceus 
series. Although obviously rare, these species were described with suf- 
ficiently distinctive characters to warrant their retention. With continued 
search, it is possible that they may be reisolated. 

Aspergillus delacroixii (Sacc.) Thorn and Church, in The 
Aspergilli, p. 190. 192G. 

Synonym: S. ochracea Delacroix, in Bull. Soc. Myc. France 7: 109, 
PI. VII, fig. f. 1891. (Delacroix failed to recognize the 
previous use of the specific name.) 
S. delacroixii Sacc, in Sacc. 10: 527. 

This species is reported as having conidiophores pale yellow and rough, 
500 to 1000m in length; vesicle globose, thick-walled, punctate, yellow; 
sterigmata in two series, primary 39/x by 12m, secondary (from Delacroix's 
figures) about 8 to 10m by 2 to 3m; conidia globose, finely roughened, 7 to 8m 
in diameter. The yellow and roughened conidiophores ally this species with 
the A. ochraceus groups hence would justify the description, if a form with 
such large conidia should be found again. Otherwise, it is possible that 
some old material of a strain of A. oryzae might have furnished the type. 

Aspergillus butyracea (Bainier) n. comb. 

Synonym: S. butyracea Bainier, in Bull. Soc. Bot. France 27: 29. 1880. 

Specimen attributed to Bainier in C. Roumeguere's Fungi 

Gallici Exsiccati No. 995. 

This is described as a large-spored strain belonging in this group. The 
specimen showed a black Aspergillus, as well as an ochraceous form with 
spores up to about 6m and rough. Colonies butter yellow, including 
conidiophores, heads, and conidia; conidiophores yellow, in mounts finely 
punctate or pitted, 13 to 16m in diameter; sterigmata primary up to 25m, 
secondary 10 to 12m in length; conidia described as smooth 5.2m, but those 
found in the material were rough and up to 6.3m- It is entirely possible 
that this species may be found again, but is has not been reported since 
Bainier described it. 

A. penicillopsis (Henngs.) Racib., P. Hennings Synonym Stilbothamnium 
penicUlopsis P. Henn. and E. Nym. described in Fungi Monsunenses 
(Warburg. O. Monsunia, Bd. I, p. 37. 1900. Leipzig); exsiccati of 
type in Pathological Collections, U. S. Dept. Agr. Bureau of Plant In- 
dustry, as: Raciborski no. 87, in Crypt. Parasiticae Java. 

This material shows an Aspergillus with the color and general appearance 
of .4. ochraceus but of gigantic proportions. Measurements as follows: 



THE ASPERGILLUS OCHRACEUS GROUP 283 

Conidiophore 50 to 70m in diameter, 10 mm. or more long with walls 7 to 
12/x thick. Surface ragged and more or less pitted. Vesicle up to 175m 
in diameter with walls 7m thick, marked with a deep pit for each sterigma. 
Sterigmata, primary 50 to 90m, at times 120 by 8 to 10m at the outer end, 
sometimes with one cross wall; secondary 15 to 25m by 3 to 4m, clustered on 
the apex of the primary. Occasionally with a sterile cell interposed between 
secondaries and primaries. Conidia 8 to 12m by 5 to 8m, elliptical, pitted, 
yellowish. 

Gigantic forms such as this occasionally appear under field conditions, 
hence reach fungus herbaria. Whether these are really different from some 
of the usual species can only be determined by collection and laboratory 
cultivation. Until such study has been made, such forms as A. penicil- 
lopsis must be questioned. 

Aspergillus ostianus Wehmer, in Bot. Centralb. 80: 449-461. 1899; also 
Monogr. pps. 117-119, Taf. II, No. 1. 1899-1901. 

Colonies upon Czapek's solution agar growing fairly well, producing a 
surface growth of crowded conidiophores and conidial heads in yellowish 
to ochraceous shades, passing to shades of cinnamon in very old cultures 
with reddish-brown colors in reverse (Wehmer reported rusty-yellow, pale 
to deep brownish-yellow to cinnamon). Conidial heads globose, up to 
200m in diameter. Conidiophores yellow, coarsely roughened, mostly 500 
to 700m long in crowded areas, becoming 1 to 2 mm. in margins of old 
colonies and usually 7 to 10m in diameter, with walls heavy, up to 2 or 2.5m 
in thickness. Vesicles commonly globose, about 40m m diameter, oc- 
casionally much larger up to 70m, hi growing heads thin-walled, colorless, 
crushing easily, leaving the funnel-like yellow tip of the conidiophore open. 
Sterigmata in two series: primary from 15 to 20m long in smaller heads 
to 35m by about 8m at the tip in large heads, secondary 10 to 13m by about 
3m- Conidia commonly 3 to 4m or even 5m in long axis, varying from pyri- 
forrrj to elliptical, or at times subglobose, rough. Sclerotia occasionally 
present but not conspicuous. 

This diagnosis was based upon Wehmer 's description, and is reasonably 
well represented by strain NRRL No. 420 (Thorn No. 4724.35) received 
from Raistrick in 1924 and reported by him to have come from Westerdijk 
as Wehmer 's original strain. With conspicuous appearances suggesting 
relationship to the A. ochraceus group, the shape and ultimate color of the 
ripe spores suggest a border line position between A. ochraceus and A. 
tamarii. 

Aspergillus sparsus Raper and Thorn, in Mycologia 36: 572-574, 

fig. 6. 1944. 

Colonies upon Czapek's solution agar at room temperature spreading 
broadly, dull grayish-brown in color, at first largely submerged, but later 



284 



A MANUAL OF THE ASPERGILLI 



developing limited aerial growth, giving rise to widely scattered erect 
conidial structures (fig. 76 A) characterized by dull greenish-tan heads not 
affecting the color of the colony as a whole; reverse in brown shades; odor 
Colonies upon hay infusion agar spreading broadly, almost wholly 



none. 



submerged, giving rise to scattered but conspicuous conidial structures 
(fig. 76 B), often in definite concentric zones; heads globose, radiate, in olive- 




Fig. 76. Aspergillus sparsus: A and B, Colonies on malt extract and hay infusion 
agars, respectively, 2 weeks; note scattered heads in B and almost complete absence 
in A. C, Conidial heads, X 18. D, Single conidial head showing globose vesicle and 
roughened conidiophore, X 425. 

buff shades (Ridgway, PI. XL). Colonies upon malt extract agar spreading 
irregularly, floccose, 1 to 2 mm. deep, cream-buff in color ; conidial structures 
very few in number; heads globose, radiate, dull olive-yellow in color 
(Ridgway, PI. XL). Conidial structures generally few in number, never 
abundant, erect, arising from a submerged mycelium; heads typically glo- 
bose becoming more or less radiate in age (fig. 76 C), mostly 200 to 250m 



THE ASPERGILLUS OCHRACEUS GROUP 285 

in diameter, occasionally as much as 500m, pale olive-buff to olive-buff 
(Ridgway, PI. XL) upon Czapek and hay infusion agars to pale olivine or 
olivine (Ridgway, PI. XXXII) upon malt extract agar. Conidiophores 
straight, mostly 1 to \\ mm. in length by 10 to 12m in diameter, approxi- 
mately uniform in diameter throughout, wall 1.2 to 1.5m in thickness, 
conspicuously echinulate, typically arising from a foot cell enmeshed in a 
network of "feeder" hyphae, often tapering abruptly in the region im- 
mediately beneath the vesicle. Vesicle comparatively thin-walled, globose 
(fig. 76 D), mostly 40 to 50m in diameter, occasionally larger or smaller, 
bearing sterigmata over the entire surface. Sterigmata in two series, 
primaries crowded, comparatively short and stout, commonly 8 to 10m by 
3 to 5m, secondaries 6 to 8m by 2.5 to 3.5m- Conidia pale yellowish in mass, 
individually showing slight coloration, subglobose to slightly elliptical, 
very finely roughened, mostly 3 to 3.5m in long axis. 

Type culture NRRL No. 1933 was isolated in February 1943, from soil 
collected in La Lima, Honduras, by Dr. L. A. Llnderkofler. A second strain 
which duplicates the type almost exactly was subsequently isolated from 
soil collected in Bixar County, Texas, by Sister Mary Clare of Our Lady of 
the Lake College, San Antonio, Texas. 

The correct position of this species within the genus Aspergillus is open to 
question. The presence of a colored, coarsely roughened conidiophore indi- 
cates close relationship with Aspergillus ochraceas. This is likewise sup- 
ported by the globose vesicle and head, although these characters are typical 
of other groups as well. The scarcity of fruiting structures upon all media, 
and more particularly the greenish tint of the spore masses, however, tend 
to set it apart from the common representatives of this great group. The 
general habit of the colonies together with the paucity of conidial structures 
is strongly suggestive of Aspergillus alliaceus, but this latter species does not 
show any trace of greenish color in its conidial heads; it does possess smooth, 
colorless conidiophores, and upon ordinary culture media regularly produces 
an abundance of black sclerotia which very often dominates and charac- 
terizes the culture. In the color and character of its conidial heads A. 
sparsus is somewhat suggestive of George Smith's new species, A. avenaceus 
(Trans. Bui. Mycol. Soc. 25: 24-27, PI. I. 1943), but it differs from this, 
as it does from A. alliaceus, in possessing rough conidiophores and in its 
failure to produce sclerotia. Until additional related forms are isolated, 
we believe it best to consider this species as a member of the A . ochraceus 
group, realizing that it does not entirely fit this placement as the group has 
hitherto been considered. 

Occurrence and Economic Importance 

Members of the A . ochraceus group are widely distributed in nature and 
can be obtained from a variety of sources. They are especially common 



286 A MANUAL OF THE ASPERGILLI 

in soils and have been isolated from samples collected in many parts of the 
world. Within the group, strains approximating the species A. ochraceus 
are by far the most abundant, although heavy sclerotium-producing strains 
approximating A. quercinus are not infrequently encountered. They are a 
common component of the microflora of decaying vegetation, but there is 
little evidence that they play a very active role in processes of decomposi- 
tion. 

Huber (1933) found a member of the group, A. sclerotiorum, capable of 
rotting apples and pears. Members of the group are commonly found in 
musty or moldy cereal grains, but are not as characteristic of this sub- 
stratum as is Aspergillus candidus and members of the A. glaucus group. 
In at least one instance A. ochraceus was reported as a human pathogen 
(Ceni, 1905). 

In the Orient, A. ochraceus and allied species constitute a portion of the 
mold flora characteristically found on "Katsuobushi" and other fermented 
preparations made from fish (Yukawa, 191 1). Aspergillus melleus Yukawa 
was isolated from such material. Because of the mixture of forms present, 
including members of the A. glaucus and A . flavus-oryzae groups, it is prob- 
ably incorrect to say that any particular species or group of species is 
responsible for this fermentation. (See also Hanzawa, 1911). 

A. ochraceus has been used to bring about desired changes in the flavor 
of coffee, and its use is covered by U. S. Patent No. 1,313,209. Samples 
of the fermenting coffee showed the organism used to be a strain of A. 
ochraceus indistinguishable from Wilhelm's species. Whereas A. niger, 
A . tamarii, and A . flavus were also capable of developing in the fermenting 
coffee, A. ochraceus alone of the species tried gave a satisfactory flavor. 

As a whole, the A. ochraceus group constitutes a very abundant, but little 
studied, group of molds. Whether the scarcity of published reports re- 
garding biochemical activities indicates an absence of such, or whether it 
merely reflects a limited amount of investigation, can at present only be 
guessed. 



PART III 

REFERENCE MATERIAL 



Chapter XXII 
TOPICAL BIBLIOGRAPHY 

In preparing this manual, the writers have considered it inadvisable to 
attempt to discuss the biochemical activities of the Aspergilli, since this 
subject is book-length in itself. Furthermore, the aim of the manual is to 
provide the mycologist and microbiologist with a means of identifying and 
interpreting Aspergilli as they are isolated from nature and to furnish the 
chemist, working with molds, with a guide which will enable him to main- 
tain industrially important cultures in an optimum condition. Neverthe- 
less, because of the increasing importance of the Aspergilli as agents re- 
sponsible for industrial fermentations, as subjects for physiological and 
biochemical investigations, and now as possible sources of various anti- 
biotics, it has seemed advisable to present a topical bibliography dealing 
with these and related subjects. We have not attempted to present a 
complete bibliography, but rather to present a sufficient number of refer- 
ences to provide the investigator and student with an entree to the literature 
of particular fields. An attempt has been made to choose the more im- 
portant papers for citation. Believing that more recent contributions will 
generally be of the greatest interest and value to the user, this topical 
bibliography is presented in chronological, rather than alphabetical, order. 
A list of subjects under which references are presented follows. 

CONTENTS OF TOPICAL BIBLIOGRAPHY 

Acid Production by Aspergilli 290 

Citric Acid 290 

Fumaric Acid 293 

Gallic Acid and Tannin Fermentation 294 

General Papers and Reviews 294 

Gluconic Acid 295 

Itaconic Acid 297 

Kojic Acid 297 

Oxalic Acid 298 

Miscellaneous Acids 299 

Antibiotics and Toxins 300 

Chemistry of Mold Tissue 301 

Enzyme Production by Aspergilli 302 

Aspergillus flavus-oryzae group 302 

Aspergillus niger group 304 

Aspergilli, General 305 

Fat Production by Aspergilli 306 

Pathogenicity of the Aspergilli 307 

Physiology of the Aspergilli 310 

289 



290 A MANUAL OF THE ASPERGILLI 

Pigments and Coloring Substances 312 

Soil Tests for Mineral Deficiencies 313 

Variation in the Aspergilli 314 

Vitamins and Growth Substances 316 

Miscellaneous Products 316 

Alcohol 316 

Chitin 317 

Ergosterol 317 

Fluorescein 317 

Gums 317 

Hydroxylamine 317 

Mannitol 317 

Polysaccharides 31S 

Ochracin 318 

Terrein 318 

Acid Production 

Citric Acid 

Wehmer, C. 1893. Note sur la fermentation citrique. Bull. Soc. Chim. France 9: 

728-730. 
Wehmer, C. 1893. Preparation d'acide citrique de synthese, par la fermentation 

du glucose. Compt. Rend. 117: 332. 
Wehmer, C. 1894. Process of making citric acid. U. S. Patent 515,033. Febru- 
ary 20. 
Wehmer, C. 1897. Uber Zwei weitere freie Citronensaure bildende Pilze. Chem. 

Ztg. 21: 1022-1023. 
Wehmer, C. 1912. Uber Citronsauergarung. Chem. Ztg. 36: 1106-1107. 
Currie, J. N. 1917. The citric acid fermentation of Aspergillus niger. Jour. Biol. 

Chem. 31: 15-37, pis. 2, figs. 5. 
Molliard, M. 1919. Production d'acide citrique par le Sterigmatocystis nigra. 

Compt. Rend. Acad. Sci. (Paris) 168: 360-363. 
Butkewitsch, W. 1923. Uber die "Citronensauregarung." Biochem. Ztschr. 142: 

195-211. 
Molliard, M. 1924. Influence de la nature des sucres sur la formation d'acides 

organiques par le Sterigmatocystis nigra en milieu desequilibre. Compt. 

Rend. Soc. Biol. (Paris) 90: 1395-1397. 
Wehmer, C. 1925. Bildung von citronensaure aus glykonsaure durch pilze. Ber. 

Deut. Chem. Gesell. 58: 2616-2619. 
Bernhauer, K. 1926. liber die Saurebildung durch Aspergillus niger. III. Die 

Bedingungen der Zitronensaurebildung. Biochem. Zeitschr. 172: 324-49. 
Henneberg, W. 1926. Handbuch der garungsbakteriologie, 2 v., illus. Berlin. 
Amelung, H. 1927. Beitrage zur Saurebildung durch Aspergillus niger. Zeit. 

Physiol. Chem. 166: 161. 
Challenger, F., Subramaniam, V. and Walker, T. K. 1927. The mechanism of 

the formation of citric and oxalic acids from sugar by Aspergillus 'niger. 

Jour. Chem. Soc. (London) 200-208. 
Kostytschev, S. and Tchesnokov, W. 1927. Bildung von Zitronensaure und 

Oxalsaure durch Aspergillus niger. Planta, Abt. E. Zeits. Wiss. Biol. 4: 

181-200. 



TOPICAL BIBLIOGRAPHY 291 

Walker, Th. K., Subramaniam, V. and Challenger, F. 1927. The mechanism of 
the formation of citric and oxalic acids from sugars by Aspergillus niger. 
II. Jour. Chem. Soc, London, 3044-54. 

Bernhauer, K. 1928. Zum Chemismus der Citronensaurebildung durch Pilze. 

I. Die Saurebildung aus verschiedenen Kohlenstoffverbindungen. Biochem. 
Zeitschr. 197: 309-26. 

Bernhauer, K. 1928. Zum Chemismus der Citronensaurebildung durch Pilze. 

II. Die Citronsaurebildung aus Gluconsaure. Biochem. Zeitschr. 197: 
327-42. 

Bernhauer, K. and Schon, K. 1928. Zum Chemismus der Citronensaurebildung 
Durch Pilze. III. Mitteilung: Uber die Hypothesen Citronensaurebildung 
und das Auftreten von Acetaldehyd in den Pilzkulturen. Biochem. Ztschr. 
202: 164-179. 

Bernhauer, K. 1928. Beitrage zur Enzymachemie der durch Aspergillus niger 
Bewirkten Saurebildungsvorgange, I. Ztschr. Physiol. Chem. 177: 86-106. 

Bernhauer, K. 1928. Uber die Saurebildung bei Aspergillus niger. IV. Die 
Bedeutung der Myzelentwicklung fur die Saurebildung. Biochem. Zeitschr. 
197: 287-308. 

Fernbach, A. and»Yuill, J. L. 1928. Improvement in and relating to the processes 
for the production of citric acid. U. S. Pat. 1,691,965 and 1,691,966. 

Filosfov, M. S. and Malinovskii, V. E. 1928. O limonno-kislom drozenii. 
Nauchnuie Zapiski 5: 235-239. 

Szucs, J. 1928. Process for producing citric acid by means of fermentation. 
Canad. Pat. 251,180. 1925, and U. S. Pat. 1,679,186. 

Cahn, F. J. 1929. Perfectionnements a la fabrication de l'acide citrique par 
fermentation. Fr. Pat . 675,236 and 675,237. 

Challenger, F. 1929. The production of citric acid by fermentation process. 
Ind. Chem. 5: 181-184. 

Amelung, H. 1930. Wachstum und Saurebildung von Aspergillus niger unter 
Wasser. Chem. Ztg. 54: 118. 

Chrzaszcz, T. and Tiukow, D. 1930. Biochemische Umbildungen der Essigsaure 
durch Schimmelpilze und liber den Chemismus der Citronensaurebildung. 
Biochem. Zeit. 229: 343. 

Bernhauer, K., SiEBENauGER, H. and Tschinkel, H. 1931. Zum Chemismus der 
Citronensaurebildung durch Pilze. IV. tlber die Umwandlung der Zuck- 
ersaure. Biochem. Ztschr. 230: 466-474. 

Bernhauer, K. and SiEBENauGER, H. 1931. Zum Chemismus der Citronensaure- 
bildung durch Pilze. V. Die Citronensaurebildung aus Essigsaure. Bio- 
chem. Zeit. 240: 232-44. 

Taylor, J. N. and Boyd, R. G. 1931. United States citric acid industry now self- 
sufficient. U. S. Dept. Com., Bur. Foreign and Dom. Com., Com. Rpt. 4: 
227-228. 

Bernhauer, K. 1932. Die oxydative Garungen. Verlag Julius Springer, Berlin. 

May, O. E. and Herrick, H. T. 1932. Production of organic acids from carbo- 
hydrates by fermentation. U. S. Dept. Agr. Circular 216. 

Porges, N. 1932. Citric Acid Production by Aspergillus niger. Am. Jour. Bot. 19: 
559-567. 

Sakaguchi, K. and Yamaya, J. 1932. Production of citric acid by molds. III. 
Japanese black molds. Jour. Agr. Chem. Soc. Japan 8: 489-97. 

Kostytschev, S. and Berg, V. 1933. Conditions of Biochemical Production of 
Citric Acid. Bull. State Inst. Agr. Microbiol. USSR 5: 8-27. 



292 A MANUAL OF THE ASPERGILLI 

Butkewitsch, W. S., Menzschinskaja, E. W. and Trofimova, E. I. 1934. Zur 
Biochemischen Herkunft von Citronen-und Oxalsaure. I. Uber die Bildung 
von Citronensaure aus Essigsaure. Biochem. Zeit. 272: 290. II. Die 
Mycelsubstanzen als ein Quelle der Saurebildung. Biochem. Zeit. 272: 364. 

Doelger, W. P. and Prescott, S. C. 1934. Citric Acid Fermentation. Ind. Eng. 
Chem. 26: 1142-49. 

Sotnikov, E. 1934. Production of citric acid by Aspergillus niger. I. Production 
of citric acid in unchanged solutions. Compt. Rend. Acad. Sci. USSR 3(4): 

273. 
Sotnikov, E. 1934. Production of citric acid by A spergillus niger. II. Production 
of citric acid in changed solutions. Compt. Rend. Acad. Sci. USSR 3(4): 

279-83. 
Sotnikov, E. 1934. Production of citric acid by Aspergillus niger. III. Produc- 
tion of citric acid in often-changed solutions. Compt. Rend. Acad. Sci. 

USSR 3(7): 544-7. 
Zubkova-Gitler, S. R. and Zalts, R. M. 1934. Preparation of Citric Acid from 

Wood-Pulp Hydrolysates. Lesokhimicheskaya Prom. 3: 4-7. 
Butkewitsch, V. S. and Gaewskaya, M. S. 1935. Yield of citric acid from sugar 

as a basis for estimating the mechanism of its formation from the latter. 

Compt. Rend. Acad. Sci. U.S.S.R. (N.S.) 3: 405-408. Abstracted in Chem. 

Abstr. 
Cahn, F. J. 1935. Citric acid fermentation on solid materials. Ind. Eng. and 

Chem. 27: 201-204. 

Chrzaszcz, T. and Peyros, E. 1935. Optimale Bedingungen der Citronensaurean- 
haufung, sowie einiger Beobachtung zu Theorie der Citronsaurebildung. 
Biochem. Ztschr. 280: 325-336. 

Giordani, M. 1935. Fermentazione citrica. La Chimica e 1'Industria 17: 77-81. 

Gudlet, M. A. 1935. Gasregime of Aspergillus niger as related to citric acid forma- 
tion. Proc. Inst. Sci. Res. Food Ind. (Leningrad) 3, No. 1: 45-69. Ab- 
stracted in Chem. Abst. 

Ivanov, N. N. 1935. Biochemical production of citric acid. Proc. Inst. Sci. Re- 
'search Ind. (Leningrad) 3, No. 1, 3-4. Practical and theoretical studies. 

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Bernhauer, K. and Iglauer, A. 1936. Uber die Saurebildung aus Zucker durch 

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Nakazawa, R., Takeda, Y and Nakano, M. 1937. Citric Acid Manufacture by 
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L'vov, S. and Toupizina, G. M. 1938. Effect of sodium fluoride on the formation 
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Zender, J. 1938. Production of Citric Acid. U. S. Patent 2,121,063. June 21. 

Bernhauer, K. 1939. Garungschemisches Praktikum. Zweite Ouflage. pp. 317. 
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Zhuravskii, G. I. 1939. The Gas Exchange in Aspergillus niger during the Forma- 
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Berhauer, K., Knobloch, H. and Zippelius, O. 1940. Uber die Saurebildung 
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Das Gupta, G. C, Saha, K. C. and Guha, B. C. 1940. The fermentative produc- 
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Mel'nikova, A. A. and Trofimova, E. I. 1940. Activation of the process of citric 
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Prescott, S. C. and Dunn, C. D. 1940. Industrial Microbiology. Chapt. XXV: 
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Bernhauer, K., Iglauer, A., and Knobloch, H. 1941. Uber die Saurebildung 
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Karow, Edward Otto. 1942. The Production of Citric Acid in Submerged Cul- 
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Sztics, Joseph. 1944. Method of producing citric acid by fermentation. U. S. 
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Fumaric Acid 

Ehrlich, F. 1911. Uber die Bildung von Fumarsaure durch Schimmelpilze. Ber. 
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294 A MANUAL OF THE ASPERGILLI 

Thies, W. 1931. Decomposition of salts of organic acids by the mold fungus 

Aspergillus fumaricus. Ber. 64B: 214-8. 
Chrzaszcz, T. and Zakoronorny, M. 1933. Biochemical Transformation of Sugar 

by Fungi. The Transformation of Fumaric Acid, the Accumulation of 

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Gallic Acid and Tannin Fermentation 

Scheele, K. 1787. tTber das wesentliche gallapfelsalz. Crell's Chem. Ann. 1: 

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184, illus. 
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Chi-Laing Kuo. 1939. Researches on Fermentation for the Production of Gallic 

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Kwang-Chur Hsieh. 1939. Researches on Fermentation for the Production of 

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Wen-Teh Wei. 1939. Researches on Fermentation for the Production of Gallic 

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General — Acid Production 

Elfving, F. 1918-19. tJber die Bildung organischer Sauren durch Aspergillus 
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Falck, R. and Van Beyma, Thoe Kingma. 1924. Methodisches und Prinzipielles 

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Herrick, H. T. and May, O. E. 1929. Molds and Chemical Manufacture. Ind. 

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May, O. E. and Herrick, H T. 1930. Some minor industrial fermentations. Ind. 

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May, O. E. and Herrick, H. T. 1932. Production of organic acids from carbo- 
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May, O. E. and Herrick, H. T. 1934. Some practical and theoretical aspects of 

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Gluconic Acid 

Molliard, M. 1922. Sur une nouvelle fermentation acide produite par le Sterig- 

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Butkeayitsch, W. 1924. Uber die Bildung der Glucon-und Citronensaure in den 

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Muller, D. 1928. Studien uber ein neues Enzym Glykoseoxydase. I. Biochem. 

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296 A MANUAL OF THE ASPERGILLI 

Schreyer, R. 1931. Comparative studies on the formation of gluconic acid by 

molds. Biochem. Zeit. 240: 295-325. A. wentii and A. luchuensis. 
Sakaguchi, K. 1932. The production of acids and alcohol by Aspergillus. IV. The 

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Japan 8: 264-265. 
Angeletti, A. and Ponte, D. 1933. Arione de al cuni funghi zu soluzioni di 

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Biochem. Zeit. 266: 337-351. 
May, O. E., Herrick, H. T., Moyer, A. J. and Wells, P. A. 1934. Gluconic acid 

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Bolcato, V. 1935. La reazione del mezzo e l'attivita dei feltri communi e pre- 

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Herrick, H. T., Hellbach, R. and May, O. E. 1935. Apparatus for the appli- 
cation of submerged mold fermentations under pressure. Ind. Eng. Chem. 

27: 681-683. 
Kardo-Syssojeva, E. 1935. Biochemical Production of Calcium Gluconate. Prod. 

Inst. Sci. Res. 3: 83-94. 

Moyer, A. J., Wells, P. A., Stubbs, J. J., Herrick, H. T. and May, O. E. 1937. 
Gluconic acid production. Development of inoculum and composition of 
fermentation solution for gluconic acid production by submerged mold 
growths under increased air pressure. Ind. Eng. Chem. 29: 777-781. 

Wells, P. A., Lynch, D F. J., Herrick, H. T. and May, O. E. 1937. Trans- 
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Wells, P. A., Moyer, A. J., Stubbs, J. J., Herrick, H. T. and May, O. E. 1937. 
Gluconic acid production. Effect of pressure, air flow, and agitation on 
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Gastrock, E. A., Porges, N., Wells, P. A. and Moyer, A. J. 1938. Gluconic 
acid production on pilot plant scale: effect of variables on production by 
submerged mold growth. Ind. Eng. Chem. 30: 782-789. 

Lvoff, S. and Loupizina, G. M. 1938. Effect of sodium fluoride on the formation 
of citric and gluconic acid by the fungus Aspergillus niger. Compt. Rend. 
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Moyer, A. J., Umberger, E. J. and Stubbs, J. J. 1940. Fermentation of Concen- 
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Porges, N., Clark, T. F. and Gastrock, E. A. 1940. Gluconic Acid Production: 
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Knobloch, H. and Mayer, H. 1941. Uber die bildung von d-glucon-, d-mannon- 
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Porges, N., Clark, T. F. and Aronovsky, S. I. 1941. Gluconic Acid Production: 
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Carrero, Jaime Gonzalez. 1942. Obtaining Gluconic Acid by the Fermentation 
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Itaconic Acid 

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Kinoshita, K. 1931. tlber die produktion von itaconsaure und mannit durch 
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Kojic Acid 

Saito, K. 1907. tlber die Saurebildung bei A spergillus oryzae. Bot. Mag. (Tokyo) 

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Rayman, M. M. 1941. Production of chemicals by the fermentation of the acid 
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Oxalic Acid 

Duclaux, E. 1889. Sur la nutrition intracellulaire. Ann. Inst. Pasteur 3: 97- 

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Miscellaneous Acids 

Bernhauer, K., Bocki, X. and Sichenauger, H. 1932. The formation of acid 

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300 A MANUAL OF THE ASPERGILLI 

Antibiotics and Toxins 

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310 A MANUAL OF THE ASPERGILLI 

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312 A MANUAL OF THE ASPERGILLI 

Steinberg, R. A. 1936. Relation of accessory growth substances to heavy metals, 

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Steinberg, R. A. 1938. Correlation between Biological Essentiality and Atomic 

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Pigments and Coloting Substances 

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Soil Tests for Mineral Deficiencies 

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314 A MANUAL OF THE ASPERGILLI 

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Variation 

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vegetatifs et conidiens, de V Aspergillus fumigatus Fresenius en cultures sur 

milieux dissocies et non dissocies sous 1 'influence des radiations du radium. 

Bull. Sci. Pharmacol., 34: 193-202. 
Barnes, B. 1928. Variations in Eurotium herbariorum (Wigg.) Link, induced by 

the action of high temperatures. Ann. of Bot. 42: 783-812. 
Sartory, A., Sartory, R. and Meyer, J. 1928. Contribution a l'etude biologique 

de 1' Aspergillus fumigatus Fresenius issu de souches sexuees et asexuees. 

Compt. Rend. Soc. Biol., 98: 215-21. 
Blochwitz, Adalbert. 1932. Variability und Vererbung bei Schimmelpilzen. 

Ber. Deut. Bot. Gesell. 50: 248-255. 
Galloway, L. D. 1933. The stimulation by dilute antiseptics, of sectoring in 

mould cultures. Brit. Mycol. Soc. Trans. 18(11): 161-162. Salicylanilide 

at concentration of 0.003 to 0.005 percent of the sodium salt produced 

sectoring. 
Greene, H. C. 1933. Variation in single spore cultures of Aspergillus fischeri. 

Mycologia 25: 117-138. 
Henrard, Paul. 1934. Polarite, Heredite, et Variation chez diverses especes 

d'Aspergillus. La Cellule, tome XL1II, fasc. 3: 351-424. 



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Mosseray, Raoul. 1934a. Les Aspergillus de la section niger Thorn and Church. 
La Cellule, tome XLIII, fasc. 2. 203-285. 

Mosseray, R. 1934b. Races naturelles et variations de culture chez divers Asper- 
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Kresling, E. and Stern, E. 1936-37. tJber die Wirkung von radium- und ultra- 
violetten Strahlen auf die Entwicklung, die biochemischen Eigenschaften 
und die Rassenbildung des Aspergillus niger. Zentralblatt fur Bakt. Abt. 
II, 95: 327-40. 

Sartory, A., Sartory, R. and Meyer, J. 1936. Etude de Taction du radium sur 
{'Aspergillus fumigatus Fresenius en culture sur milieux dissocies et non 
dissocies. Compt. Rend. Acad. Sci., 183: 77-79. 

Nakazawa, R. and Simo, M. 1938. Effect of irradiation on fermentative micro- 
organisms. I. Morphological and biochemical characteristics of races of 
Aspergillus niger produced by radium treatment. Jour. Agr. Chem. Soc. 
Japan 14: 895-910. 

Whelden, R. M. 1938. Changes observed in cultures of Aspergillus niger bom- 
barded as spores with low voltage cathode rays. Mycologia 30: 265-268. 

Yuill, Edward and Yuill, John L. 1938. Cladosarum olivaceum. A new hy- 
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Btjchwald, C. E. and Whelden, R. M. 1939. Stimulation of growth in Aspergillus 
niger under exposure to low velocity cathode rays. Am. Jour. Bot. 26: 778- 
784. 

Nakazawa, Ryozi and Simo, M. 1939. The action of radium and x-rays on micro- 
organisms. II. The production of acids by radium-irradiated Aspergillus 
niger. Jour. Agr. Chem. Soc. Japan 15: 547-52. 

Thom, Charles and Steinberg, Robert A. 1939. The chemical induction of 
genetic changes in fungi. Proc. Nat. Acad. Sci. 25: 329-335. 

Yuill, Edward. 1939. Two new Aspergillus mutants. Jour, of Botany, 174-175, 
pi. 618. June. 

Zahl, Paul A., Koller, L. R. and Haskins, C. P. 1939. The effects of ultra- 
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Gossop, George Harold, Yuill, Edward and Yuill, John Lewis. 1940. Hetero- 
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Simo, M. 1940. Experimentelle Untersuchungen uber die Wirkung von Radium 
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Bedingung der Citronensauregarung durch Aspergillus niger Radium rasse. 
Jour. Agr. Chem. Soc. Japan 16: 129. 

Steinberg, R. A. and Thom, Charles. 1940. Chemical induction of genetic 
changes in Aspergilli. Jour, of Heredity 31: 61-63. 

Steinberg, Robert A. and Thom, Charles. 1940. Mutations and reversions in 
reproductivity of Aspergilli with nitrite, colchicine and d-lysine. Proc. 
Nat. Acad. Sci. 26(6): 363-366. 

Whelden, Roy M. 1940. "Mutations" in Aspergillus niger bombarded by low 
voltage cathode rays. Mycologia 32: 630-643. 

Steinberg, R. A. 1942. Reversions in morphology of nitrite-induced "Mutants" 
of Aspergilli grown on amino acids. Jour. Agr. Res. 64: 645. 

Thom, Charles. 1942. Chemical induction of genetic changes in Aspergilli. Jour. 
Franklin Inst. 233:284. 



31G A MANUAL OF THE ASPERGILLI 

Vitamins and Growth Substances 

Boysen-Jensen, P. 1931. Formation of a growth regulator by Aspergillus niger. 
Biochem. Zeit. 239: 243-9. 

Schopmeyer, H. and Fulmer, E. I. 1931. The production of yeast growth stimu- 
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gillus clavatus. Jour. Bact. 22: 23-28. 

Sakamura, T. and Yanagihara, T. 1932. The production of the growth substance 
by Aspergillus niger. Proc. Imp. Acad. Tokyo 8: 397-9. 

Schopmeyer, H. 1932. Production of yeast-growth stimulants by molds on various 
media. la. St. Coll. Jour. Sci. 6: 471-2. 

Bunning, E. 1934. Growth and nitrogen assimilation in Aspergillus niger under 
the influence of growth regulators. Ber. Deut. Botan. Ges. 52: 423-44. 

Bernhauer, K. and Gorlich, B. 1936. The formation of vitamin C-like sub- 
stances by fungi and bacteria. I. Biochem. Zeit. 286: 60. 

Komarov, S. N. 1936. Methods of obtaining an antirachitic preparation from 
the waste products of citric acid manufacture from the mycelium of Asper- 
gillus niger. Proc. Sci. Inst. Vitamin Res. USSR 1(2): 162-71. 

Opfel, V. V. 1936. Chemistry and biochemistry of flavins. Vitamins and vita- 
minization. Proc. Sci. Inst. Vitamin Res. USSR 1(2): 5-52. 

Scheunert, A. and Schieblich, M. 1936. Vitamin production by Aspergillus 
oryzae. Biochem. Zeit. 286: 66-71. 

Fukumoto, J. and Shinomura, H. 1937. Formation of Vitamin C-like substance 
by molds. Jour. Agr. Chem. Soc. Japan 13: 613-20. 

Kitavin, G. S. 1939. Action of mercury salts on the formation of vitamin B 2 in 
Aspergillus niger. Biokhimiya 4: 283-94. 

Lavollay, J. and Laborey, F. 1939. Characterization of lactoflavin produced by 
Aspergillus niger v. Tgh. partially deficient in magnesium. Compt. Rend. 
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Knoblock, H. and Sellmann, R. 1941. The formation of flavin type pigments in 
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277-80. 

Eakin, R. E. and Eakin, E. A. 1942. Biosynthesis of Biotin. Science 96: 187. 
A. niger synthesizes biotin from biotin free media. 

Miscellaneous Products 
Alcohol 

Molliard, M. 1916. Catalytic role of potassium nitrate in alcoholic fermentation 

produced by S. nigra. Compt. Rend. Acad. Sci. (Paris) 163: 570-572. 
Yuill,J.L. 1928. Alcoholic fermentation by Aspergillus flavus Brefeld. Biochem. 

Jour. 22: 1504-7. 
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Vyatkin, V. 1940. New process for making alcohol from chicory. Spirto-Vodoch- 

naya Prom. 17(9): 13-14. 



TOPICAL BIBLIOGRAPHY 317 



Chitin 



Norman, A. G. and Peterson, W. H. 1932. The chemistry of mold tissue. II. 
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Ergosterol 

Pruess, L. M., Peterson, W. H. and Fred, E. B. 1932. Isolation and identifica- 
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Fluorescein 

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Gum 

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Hydroxylamine 

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Lemoigne, M. and Desveaux, R. 1936. Recherches sur la role biochimique de 
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Mannitol 

Braconnot, H. 1813. Nouvelles recherches analytiques sur les champignons pour 
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318 A MANUAL OF THE ASPERGILLI 

Polysaccharides 

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Ochracin 

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Terrein 

Raistrick, H. and Smith, G. 1935. Biochemistry of micro-organisms. XLII. 
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Chapter XXIII 
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319 



320 A MANUAL OF THE ASPERGILLI 

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White, E. C. 1940. Bactericidal filtrates from a mold culture. Science 92: 127. 
and Hill, J. H. 1943. Studies on antibacterial products formed by molds. 

I. Aspergillic acid, a product of a strain of Aspergillus flavas. Jour. Bact. 

45: 433^44. 
Wickerham, L. J. and Andreasen, A. A. 1942. The lyophil process: its use in the 

preservation of yeasts. Wallerstein Lab. Coram. 5(16): 165-169. 1 plate. 
Wiggers, Fredericus Henricus. 1780. Primitiae Florae Holsaticae. 114 pp. 

Kiliae. (Facsimile ed. 23 by W. Junk. 1925.) 
Wilhelm, K. A. 1877. Beitriige zur Kenntniss der Pilzegattung Aspergillus. In- 

aug. Diss. Strassburg pp. 66. 
Wilkins, W. H. and Harris, G. M. C. 1942. Investigation into the production of 

bacteriostatic substances by fungi. I. Preliminary examination of 100 

fungal species. Brit. Jour. Exptl. Path. 23: 166. 
Wilson, J. A. and Daub, G. 1925. Aspergillus niger, a common mold that causes 

black spots on leather. Jour. Amer. Leather Chemists Assoc'n 20: 400-405, 

figs. 12. 
Wolf, Frederick A. 1938. I. Fungal flora of Yucatan caves. Carnegie Inst. 

Wash. Pub. No. 491, p. 19-21. 
Woolley, D. M., Strong, F. M., Peterson, W. H. and Prill, E. A. 1935. Chem- 
istry of mold tissue. X. The phospholipides of Aspergillus sydowi. Jour. 

Am. Chem. Soc. 57: 2589-91. 
Yabuta, A. 1912. On Koji acid, a new organic acid formed by Aspergillus oryzae. 

Journ. Coll. Agr., Tokyo, 5: 51-8. Orig. Coram. 8th Intern. Congr. Appl. 

Chem., (Append.), 25 (1913) 45-62. 
Yamamoto, Y. 1927. Leber drei Stamme von Aspergillus icentii (Japanisch.) 

Jour. Soc. Agr. Forest., Sapporo, 19: 128-40. 
Yuill, Edward. 1939. Two new Aspergillus mutants. Jour. Bo£. (London): 

174-175, illus. 
and Yuill, John L. 1938. Cladosarum olivaceum, a new Hyphomycete. 

Brit. Mycol. Soc. Trans. 22: 194-200, illus. 
Yukawa, M. 1911. Zwei neue Aspergillusarten aus "Katsuobushi." Jour. Coll. 

Agr. Tokyo 1: 357-366. 
Zikes, H. 1922. Lber die Perithecienbildung bei A. oryzae. Centralb. f. Bakt. 

etc. 2 Abt. 56: 339-343. 



Chapter XXIV 
CHECK LIST OF SPECIES AND GENERA 



GENERIC NAMES FOUND APPLIED 
TO ASPERGILLI 

Alliospora G. Pirn, in Proc. R. I. 
Acad. 1883, and Jour. Bot. 1883, 
p. 234; also Sacc. Syll. 18, No. 
5216. 1906. A. Sapucaya rep- 
resents a monotypic genus found 
upon putrifying Lecythis Sapu- 
cajo. From the description it 
was some member of the A. niger 
group. Saccardo's diagnosis 
only seen 9 

Ascophora. Ascophora nigrans was 
vaguely assigned mostly to 
Mucors but appears to have been 
used by Raulin's group studying 
the tannic acid fermentation 
until Van Tieghem described 
A. niger 9 

Aspergillopsis Sopp, in Videnskaps- 
selskapets Skr. I Mat.-Na- 
turv. Kl. No. 11 , pp. 201-204, Taf . 
XX, fig. 149. 1912. Sopp 
probably had some members of 
the A . ust us group 9 

Aspergillopsis Spegazzini, in An. 
Mus. Nat. Buenos Aires Ser. 
3, 13: 434. 1911. The black 
spored Aspergilli (essentially the 
A. niger group) were regarded 
as dematiaceous and assigned 
to a new genus in that group. 
Spegazzini 's genus has not been 
accepted 8 

Aspergillus Micheli, in Nova Plan- 
tarumGenerap. 212, PI. 91. 1729. 
Compare Link, in Obs. p. 16, 
1809, and Corda in Icones 
Fungorum 4: 31, Tab. VII, fig. 

94. 1840 6 

Subgenus — Microaspergillus Weh- 
mer, in Monogr. 1901. (Mem- 
oires de la Societe de Physique 



et d'Histoire Naturelle de 
Geneve, Tome XXXIII, Seconde 
Parte, pp. 1-157. 1899- 

1901) 16 

Subgenus — Macroaspergillus Weh- 
mer in Monogr. 1901. The 
subgenera are not accepted 
here 16 

Cladosarum Yuill and Yuill, in 
Trans. Brit. Myc. Soc. 22: 194- 
200, Pis. 11-13. 1938. Re- 
garded as a laboratory mutant 
not encountered in nature. In 
A. niger group. See 73 

Dimargaris van Tieghem, in Ann. 
Sci. Nat. 6 Ser. 1: 54. 1875. 
The name Dimargaris sp. has 
been seen upon living cultures in 
one of the great laboratory col- 
lections. These cultures were 
obtained from dog and squirrel 
dung. The organism so labeled 
belonged to the Aspergillus 
candidus group. The genus 
Dimargaris (type D. crystalli- 
gena v. Tieghem) is regarded by 
Fitzpatrick as closely related to 
Dispira and as a synonym of 
Dispira by Zycha (Krypto- 
gamenflora der Mark Branden- 
burg. Pilze II. Mucorineae. 
Band Via: 1-264, 114 figs. 
1935). In any case, it is only 
reported as a parasite of the 
mycelia of the Mucorineae, 
hence certainly was not an As- 
pergillus 9 

Diplostephanus Langeron, in Compt. 
Rend. Soc. Biol. Paris 87: 343- 
345. 1922. This genus is pro- 
posed by Langeron to include 
ascosporic Aspergilli (Sterig- 
matocystis) with two series of 



331 



332 



A MANUAL OF THE ASPERGILLI 



sterigmata. A. nidulans Ei dam 
is named as type. The proposal 
is rejected here 8 

Emericella Berkeley and Broome, in 
Introd. Crypt. Bot. pp. 340-341, 
fig. 76. 1857. See also: Pa- 
touillard in Bui. Soc Myc. 
France 7: 43-49, pi. 4, figs. 6-12. 
1891. Based upon E. variecolor. 
Syn. A . variecolor q.v 163 

Euaspergillus Ludwig, in Lehrbuch 
der niederen Kryptogamen p. 
258. Stuttgart 1892. The 
genus was proposed to cover the 
sclerotium producing Aspergilli 
such as A. niger, A. flavus, A. 
ochraceus. The name has not 
been accepted 8 

Eurotium Link, in Obs. p. 31, Taf. 2, 
fig. 44. 1809. Link described 
Aspergillus glaucus as a conidial 
mold and Eurotium herbariorum 
for the ascosporic form, suppos- 
ing them to be different fungi. 
His genus Eurotium has been 
widely accepted, but it is re- 
jected here for reasons discussed 
elsewhere. Forms listed as 
Eurotium but having by descrip- 
tion black perithecia are 
excluded. No species with 
aspergilloid conidial structures 
has black (Dematiaceous) peri- 
thecia 7 

Inzengaea Borzi, in Jahrb. Wiss. Bot. 
(Pringsheim) 16: 450-463, PL 
19,20. (1884) 1885. Type sp.: 

I. erythrospora Borzi: Syn. 

A. variecolor q.v 165 

Mucor used by Wiggers (Wichers) 
in Primitiae Florae Holsaticae. 
1780. (Republished in facsimile 
Edition No. 23 by W. Junk in 
1925). "Mucor herbariorum 
sessilis luteus" is given as No. 

1158 7 

Rhodocephalus Corda, in Icones I: 
21. 1839, and the figures given for 
R. aureus in Icones III, Taf. 

II, fig. 33 suggest A. terreus. 
Sartorya Vuillemin, in Compt. Rend. 



Acad. Sci. (Paris) 184: 136-317. 
1927. Inadequately described 
as an ascosporic phase of A. 
fumigatus. It was not distribu- 
ted in culture 9 

Sterigmalocystis Cramer, in Viertel- 
jahresschrift der Naturforchun- 
gen Gesellschaft, Zurich 4: 325. 
1859. Type: A. antacustica 
Cramer. Cramer proposed to 
include in his new genus all 
aspergilloid species showing 
both primary and secondary 
sterigmata. S. nigra (A. niger) 
was the organism under discus- 
sion, hence the type species. 
Many mycologists have agreed 
with Cramer; others, such as 
Fischer, Wehmer, and Thorn, 
have rejected Sterigmato- 
cystis 8 

CHECK LIST OF NAMED SPECIES 

S. acini-uvae Caballero, in Boletin 
R. Soc. Esp. Hist. Nat. 28: 429. 

1928 229 

Syn. A. carbonarius fide Blochwitz, 
or A. pulchellus fide Mos- 
seray 229 

S. aerea Bainier, in Bui. Soc. Bot. 
France 28: 78. 1881. A.wentii 
group 249 

A. africanus Dur. et. M., in Fl. Alg., 
LP. 342. 1849. Not an Asper- 
gillus. 

A. ageni This name is cited by 
Lindt, in Arch. Exp. Path. 
Pharm. 25: 265. 1889, as taken 
from Saccardo's Sylloge. 
Search for this reference leads to 
the conclusion that in this cita- 
tion A. H ageni was made to read 
A. ageni. 

S. alba Bainier, in Bui. Soc. Bot. 
France 27: 30. 1880. Some 
member of A. candidus group. . 211 

>S. alba-cyanogena Biourge, listed 
among cultures in the Biourge 
Collection. 1939. 

S. alba-lutea Biourge, n. m., in MS. 
p. 9, among the niveus series. 



CHECK LIST OF SPECIES AND GENERA 



333 



A. alba-roseus attached to a culture 
in the Bainier Collection and by 
inference in the Biourge Collec- 
tion as near A. niveus Blochwitz. 
Thorn's No. 4640.490. 

S. alba-sclerotifera Biourge, nomen 
nudum, listed in his MS. as 
related to A. okazakii. It 
produces a violet or blue color 
in reverse. 

S. alba-sulphurea Biourge, nomen 
nudum, listed in MS. as near 
A. okazakii. It produces a blue 
or violet reverse. 

A. albidus Eichelbaum, in Verhand- 
lungen d Naturw. Ver. Hamburg 
3 Folge XIV; 35. 1906. 
Syn. E. albidum Sacc, Syll. 22: 
1254. 1913. 

A. albidus Speg. A culture under 
this label distributed by Biourge 
represents a strain of A. niveo- 
glaucus q.v 135 

S. albo-lutea Bainier, in Bull. Soc. 
Bot. France 27: 30. 1880. This 
was a small pale yellow form, but 
no further data were given and 
it has not since been identified. 
Apparently close to A. carneus. 

S. albo-lutea Sartory, Sartory, and 
Meyer, cited by Blochwitz, in 
Ann. My col. 31: 73. 1933. 
Some member of the A . candidus 
group 211 

A. albo-marginatus Biourge, in MS. 
p. 2, as change of name from 
Penicillium albo -marginatum, 
figured only in the Penicillium 
Monograph in La Cellule tome 
XXXIII. 1 fasc. fig. 129. It 
was some member of the A. 
restrictus series, received and 
studied as Thorn No. 4733.134a. 
Near A . gracilis in the A . glaucus 
group 139 

S. albo-rosea Sartory, Sartory, and 
Meyer, in Ann. Mycol. 28: 358- 
359, PI. Ill, figs. 1-6. 1930. 
Compare S. albo-rosea in 
Blochwitz Ann. Mycol. 31: 75. 
1933. A . carneus series 202 



A. albus Wilhelm, Inaug. Diss. 
Strassburg, p. 69. 1877. Prob- 
ably A. albus Haller, S. alba 
Sacc . , Monilia alba Persoon . In 
A . candidus group 211 

A. alliaceus Thorn and Church, The 
Aspergilli, p. 163. 1926. See 
A . wentii group 244 

A. alternatus Berkeley, in Ann. Nat. 
Hist., Ser. 1, Vol. 1: 262. 1838. 
Not an Aspergillus. 

A. alutaceus B. and C, in Grevillea 
3: No. 25, p. 108. 1875. In the 
A. ochraceus group, unidenti- 
fiable to strain or series 281 

S. ambari (alternative spelling, 
ambaris) Beauregard, in Ann. de 
Micrographie 10: 255-278, 1 
plate. 1898. Cited without 
description in Compt. Rend. 
Hebdom. des Seances de la Soc. 
de Biol. 28: Mai (1898). A. 
vesicolor group 192 

A. amoenus Roburg, in Hedwigia 
70: 138-9. 1930. Distributed 
by the Centraal -bureau voor 
Schimmelcultures, Baarn. Re- 
ceived by us on November 20, 
1933, and determined as a 
member of the A. gracilis series, 
but the description allies it with 
A. versicolor. 

A. amstelodami (Mangin) Thorn and 
Church, in The Aspergilli, p. 
113. 1926; also Thorn and 
Raper, U. S. D. A. Misc. Publ. 

426, pp. 22-26. 1941 122 

Syn. E. amstelodami Mangin, in 
Sci. Nat. Bot. Ser. 9, 10: 360- 
361. 1909. A. glaucus group.... 122 
Incorrectly spelled A . amsterodami 
in Nakazawa, Jour. Agr. Chem. 
Soc. Japan 10(2): 1934. 
Syn. E. repens var. amstelodami 
Vuill., in Soc. Mycol. de France, 
Bui. Trimest. 36: 131. 1920. 
var. alophote Duche, cited by 
Dodge in his Med. Mycol. p. 630. 
1935. Ascospore lacks the crest 
of the species. 



334 



A MANUAL OF THE ASPERGILLI 



A. anomalus Mosseray, in La Cellule 
XLIII: 248-249, pi. 4, figs. 107- 
112. 1934. Member of A . niger 
group 234 

S. antacustica Cramer, in Vrtljschr. 
Naturf. Gesell. Zurich, Jahrg. 
4, Heft. 4: 325. 1859. In A. 

niger group 239 

Syn. A. phoenicis in A. niger 
group 222 

A. archaeoflavus Blochwitz, in Ann. 
My col. 31(1/2): 73-83. 1933. 
The type culture (Thorn No. 250- 
5346) was too close to A. wentii 
for satisfactory separation 247 

A. archiflavipes Blochwitz, in Ann. 
Mycol. 32(1/2): 84. 1934. A 
form of A . flavipes characterized 
by deep brown to red shades of 
color in the mycelium 181 

A. argentinus Spegazzini, in Rev. 
Agr. Veter. LaPlata p. 245. 
1896. A conidial form in the 
A. glaucus group; not identifi- 
able. 

A. argillaceus was distributed but 
not described by Biourge. It 
appears to be a non-ascosporic 
strain of the A. repens series — 111 

A. atro-fuscus Mosseray, in La 
Cellule XLIII: 269-270, pi. 4, 
figs. 126-129. 1934. Member of 
A . niger group 235 

A. atropurpureus Zimmermann, in 
Centbl. Bakt. [etc.] Abt. 2, Bd. 
8, No. 5, p. 218. 1902. A. niger 
group 226 

A. atropurpureus Blochwitz, in Ann. 
Mycol. 32(1/2): 86. 1934. 
Blochwitz proposed this name to 
cover purple-brown series in A. 
niger group 225 

A. atro-ruber Estienne 

Syn. Penicillium roseo-cinnabari- 

num Biourge, in La Cellule 33: 

fasc. 11, 319-321, PI. XVII, fig. 

97. 1923'. 

Syn. Physomyces heterosporus 

Harz, fide Biourge. 
Syn. S. atro-rubra (Estienne) 
Biourge MS. 



A. atro-violaceus Mosseray, in La 
Cellule XLIII: 268-269, pi. 4, 

figs. 122-125. 1934 235 

Blochwitz in Ann. Mycol. 33: 240. 
1935, identifies this with A. 

violaceo-fuscus Gasperini 231 

A. atrovirens Karst, in Symb. 26: 28; 
Sacc. Syll. 10: 524. Inadequate- 
ly described for identification. 
A. aurantiacus Berkeley, in British 
Fungi fasc. IV: 1843. Cited 
by Montagne in Ann. Sci. Nat. 
Bot. 3. Ser., 12: 299. 1849. 
Syn. N ematogonium aurantiacum. 
S. aurea Greco, in Origine des 
Tumeurs et Mycoses Argentines, 
Buenos Aires, pp. 671-694, figs. 
418-428. 1916. Not since recog- 
nized. Some A . ochraceus strain. 
A. aureoglaucus Roburg, in Hedwigia 
70: 137. 1930. Culture some 
strain of A. repens contributed 
by Roburg. Noted as some 
member of A. glaucus group 
by Blochwitz in Ann. Mycol. 
33: 240. 1935. 
A. aureus Berkeley, in English Flora 
5: 346. 1836. The golden 
yellow, elliptical conidia re- 
ported by Berkeley suggest 
A. citrisporus but actual identi- 
fication from the description is 

impossible 219 

A. aureus Nakazawa, in Inst. Gov't. 
Res. Formosa Rept. Vol. 1. 

1907 219 

A. aureus varieties described by 
Nakazawa, Simo and Watanabe 
in Jour. Agr. Chem. Soc. Japan 
No. 144, 1936 (In Japanese). 

var . acidus 220 

var . brevis 220 

var. minor 220 

var. murinus 220 

var. pallidus 220 

comparison of figures and measure- 
ments given convince the authors 
that these varieties could not be 
safely identified by descriptive 
data. 



CHECK LIST OF SPECIES AND GENERA 



335 



S. auricoma Gueguen, in Bull. Soc. 
Mycol. France 15: 171-187, figs. 
1-48. 1899. In A. ochraceus 

group 276 

A. auriculaire Moquin-Tandon, in 
Elements Bot. Med. 2 ed., p. 466. 
1866. This name was not 
accompanied by adequate de- 
scription to separate the form 
intended from other species 
which are occasionally found in 
the ear. 
.4. ariarius Peck, in N. Y. State 
Mus. Rept. (1890), 44: 120. 
1892. A strain of A. fumigatus 

isolated from a canary 151 

A. avenaceus George Smith, in Brit. 
Mycol. Soc. Trans. 25: 24-27, 

PI. 1, figs. 1-3. 1943 246 

A. awamori Nakazawa, in Inst, of 
Gov't. Research, Formosa Rept. 
1: 1907, and 2: 1912. A member 

of the A . niger group 220 

A. awamori varieties described by . 
Nakazawa, Simo, and Watanabe 
in Jour. Agr. Chem. Soc. Japan, 
No. 144. 1936. (In Japanese). 

var. ferrugineus 220 

var. furneus 220 

var.fuscus 220 

var. minimus 220 

var. piceus 220 

No doubt the describers had a 

series of industrially significant 

strains but it is doubtful if others 

could identify them from the 

description and figures given. 

A. awamori Usami, in Myk. Cen- 

tralbl. 4: 193. 1914. Species 

recognized by Mosseray in La 

Cellule 43: 264. 1934. With A. 

awamori Nakazawa cited as a 

synonym. 

A. bainieri Mosseray, in Ann. Soc. 

Sc. Bruxelles 54, ser. B, p. 79, 

1934 233 

Syn. A. longobasidia Bainier fide 
Mosseray in La Cellule XLIII 

(2): 227. 1934 233 

A. barbae Castellani. Cited by 
Sartory, A., in Champignons 



parasites de l'homme et des 
animaux, p. 595. 1922. This 
organism was found in the beard 
of a native of Uganda and again 
from Ceylon. The only further 
information given is conidia 4 to 
5ju, globose, dark brown. 

S. basidiosepla Sartory, Sartory, and 
Meyer, in Ann. Mycol. 27: 317- 
320. PI. VII. 1929. A variant 
of the A . candidus group 211 

A. batatae Saito, in Centralb. f. Bakt. 
2, Abt. 18(1/2): 34. 1907. In 
the A . niger group 225 

A. belfanti Carbone, in Atti d. Inst. 
Bot. Univ. Pavia Serie II. Vol. 
XIV: 63, fig. 11. 1914. Prob- 
ably in A. glaucus group. 

A. (Sporodinia) bellomontii Mon- 
tagne, in Ann. Sci. Nat. Bot. Ser. 
4, V. 12: 181-182. 1859. Noted 
as related to A . maximus (Sporo- 
dinia). Not an Aspergillus. 

S. bicolor J. Ray, in Rev. Sc, Ser. 4, 
V. 8: 176-177, 193-212. Lille, 
1897. Probably in A. sydowi 
series 192" 

A. biourgei Mosseray, in La Cellule 
XLIII: 241-242, pi. 4, figs. 81- 
85. 1934. One of the A. niger 
group 234 

S. blanc-jaune Bainier, nomen 
nudum. A culture so labeled 
from the Bainier Collection rep- 
resents a diminutive member 
of the A. candidus group 211 

A. blochwitzii Biourge, nomen 
nudum, listed in Biourge MS 
(p. 3) among the A. clavatus 
group as a synonym of A. cla- 
vatus var. gigantea Blochwitz. 

A. boedijni Blochwitz, in Ann. 
Mycol. 32: 83-89. 1934. 
Syn. A. terreus var. boedijni 197 

E. bonariense Speg., in Anal, de la 
Soc. Cient. Argen. 10: 177. 
1880. Some member of the A. 
glaucus group but data are 
inadequate. 

Coremium Borzianum Saccardo. 

See A . variecolor 165 



336 



A MANUAL OF THE ASPERGILLI 



A. bouffardi Brumpt (1905). Cited 

by Castellani and Chalmers in 

Man. Trop. Dis. p. 805. 1913. 

Syn. Madurella Bouffardi (Brumpt) 

Dodge. Med. Myc. p. 685. 1935. 

A. Brodeni (Mattlet) Dodge, in 
Dodge Med. Mycol., p. 635. 

1935 170 

Syn. S. Brodeni Mattlet, in Ann. 
Soc. Beige Med. Trop. 4: 167- 
171, figs. 1, 2. 1924. Probably 
A. unguis. 
var. V ancampenhouti (Mattlet) 
Dodge, in Dodge Med. Myc, 
636. 1935. 

A. bronchialis Blumentritt, in Ber. 
Deut. Bot. Ges. 19: 442-446, pi. 
22, figs. 1-6. 1901 ; also ibid. 23: 
419-127, pi. 19, figs. 1,3, 6, 7,8, 
19, and 23. 1905. In the A. 
fumigatus group 151 

A. brunneo-fuscus See, in Les 
Malades du papier pique Paris, 
p. 29. 1919. Unidentified ex- 
cept as a member of the A. 
glaucus group. 

A. brunneo-virens Delacroix, in Bui. 
Soc. Myc. France 13: 120, with 
text figure. 1897. Unidentified 
except as a member of the A. 
glaucus group. 

A. brunneus Delacroix, in Soc. 
Mycol. Bui. France 9: 185, PI. 
XI, fig. III. 1893. Delacroix 
described this as the conidial 
stage of A. echinulatus q.v 131 

S. Buntingiana Biourge, nomen 
nudum, listed in Biourge 's MS. as 
applied to a culture of a member 
of the A. candidus group re- 
ceived from Bunting. 

A. Buntingii Mosseray, in La Cellule 
XLIII: 236-238, pi. 4, fig. 91-95. 
1934 . Member of A . niger group 

in Biourge Collection 1939 233 

A. butyracea (Bainier) n. comb 282 

Syn. S. butyracea Bainier, in Bull. 
Soc. Bot. France 27: 29. 1880. 
C. Roumeguere's Fungi Gallici 
Exsiccati No. 995 is recorded 



as Bainier's material. In the 
A . ochraceus group 282 

A. byssoides Sprengel, in Sys. Veg., 
ed. 16, V. 4: 541. 1827. A 
fungus on rotting paper with 
globose fuscous heads but not 
identifiable. 

A. cacao. This name appeared upon 
a culture in the Bainier collec- 
tion (Thorn No. 4640.397) and 
has been contributed also by 
Pribram (4777.4). The organ- 
ism is a strain of A. tam- 
arii 256 

A. caesiellus Saito, in Jour. Coll. 
Sci. Imp. Univ. Tokyo 18: 49, 
PL III, fig. 14. 1904. Re- 
garded by Neill (in Royal Soc. 
New Zealand Trans. 69: 242. 
1939) as A . restrictus G. Smith. . . 141 

A. caespitosus n. sp. Raper & Thom, 
in Mycologia 36: 563-565, fig. 3. 
1944. A member of A . nidulans 
group 166 

A. calyptratus Oudemans, in Arch. 
Neerl, II. 7: 283, pi. 13. 1902. 

See: A. fumigat us group 151 

var. italicus Ferraris, in Ann. My- 
col. 10: 294. 1912. 

S. cameleo Sartory, Sartory, and 
Meyer, in Ann. Mycol. 29: 360- 
361, PI. III. 1930. Some vari- 
ant of A . sydowi 186 

S. Candida Sacc, in Michelia 1, p. 91. 
1877. In the A. candidus 
group. 

S. candidula Bainier, in Sacc. Syll. 
4: 73. 1886. 
Syn. S. Candida Bainier, in Bui. 
Soc. Bot. France 27: 30. 1880. 
A member of the A. candidus 
group. 

A. candidus Link, in Obs. p. 16. 

1809 207 

var. thermophilus Nakazawa et al., 
in Jour. Agr. Chem. Soc. Japan 
88: 17. 1932, (in Japanese) 
cited Blochwitz Ann. Mycol. 
33: 244. 1935. 

A. capitulo pullo Haller, in Historia 
Stirpum Indigenarum Helvetiae 



CHECK LIST OF SPECIES AND GENERA 



337 



Inchoata, etc. 1768. Appar- 
ently taken from Micheli 1729. 

S. carbonaria Bainier, in Bui. Soc. 
Bot, France 27: 27-28. 1880. 
See A. carbonarius (Bainier) 
Thorn. In .4 . niger group 229 

A. carbonarius (Bainier) Thorn, in 

Jour. Agr. Res. 7: 12. 1916 229 

A. carbonarius sen aler Meis and 
Parascandalo, in Gaz. Ospedali 
16: 769-772. 1895. Cited by 
Dodge, in Med. Myc. 679, as not 
an Aspergillus. 

A. carncolus Sacc, in Michelia 1, p. 
77. 1877. And Fungi italici 
No. 18. In A. glaucus series. 

A. carneus (van Tieghem) Bloch- 
witz, in Ann. Mycol. 31(1/2): 

81. 1933 201 

Syn. Sterigmatocystis carnea van 
Tieghem, in Bull. Soc. Bot. 
France 24: 103. 1877. See also 
Saccardo Sylloge 4: 74, and 
Wehmer's Monograph (Mem. 
Soc. Phys. His. Nat. Gen. pp. 
1-157). 1899-1901. Species in 

A . terreus group 201 

var. subglobosa Blochwitz, in Ann. 

Mycol. 33: 243. 1935. 
var. opaca Blochwitz, in Ann. 
Mycol. 33: 249. 1935. 

A. carnoyi Biourge, descr. Thom and 
Raper in U. S. Dept. Agr. Misc. 
Pub. 426, p. 34-5. 1941 134 

S. castagnei Biourge, undescribed 
culture in the Biourge Collec- 
tion, listed as near A. carbo- 
narius in the A. niger group. 

A. castonea Patterson, in Bui. Torrey 
Bot. Club 27: 284. 1900. In A. 
tamarii series. 

A. cellulosae Hopffe, in Centralb. f. 
Bakt., etc., 1 abt., 83: 531-37. 
1919. 
Syn. A.fumigatus 151 

A. cervinus Massee, in Kew Misc. 
Bui. 4: 158. 1914. Neill con- 
siders A. gratioti Sartory to be a 
synonym but that is not 
probable. 



A. chevalieri (Man^in) Thom and 
Church, in The Aspergilli, p. 
Ill, 1926. 
Syn. E. chevalieri Mangin, in Ann. 
Sci. Nat, Bot. Ser. 9, 10: 361- 
362, fig. 12. 1909. In the A. 
glaucus group 118 

A. chevalieri var. intermedins Thom 
and Raper in U. S. Dept. Agr. 
Misc. Pub. 426, p. 21, fig. 8B. 
1941 121 

A. chevalieri var. rnultiascosporus 
Nakazawa, Takeda, Okada, and 
Simo, in Agr. Chem. Soc. Japan, 
Jour. 10: 135-192. 1934. Re- 
ceived from Baarn and appears 
as NRRL No. 88. A strain of 
A . chevalieri 120 

E. chilense Montagne, in Syll. Crypt. 
No. 919, Fl. Chil. VII, p. 476; 
cited by Saccardo Syll. 1, p. 27. 
In the A. glaucus group. 

S. chlorina Cooke and Massee, in 
Grevillea 18: 7. 1889. Proba- 
bly a nonascosporic strain of the 
A. glaucus group. 

A. chrysospermum Thaxter, nomen 
nudum, attached to a culture 
later shown to be A. citrisporus 
Van Hohnel, q.v. 

A. chungii Shih, in Lingnan Sci. 
Jour. 15(3): 378. 1933. In the 
A . flavus group 267 

A. churchii Mosseray, in La Cellule 
XLIII: 242-244, pi. 4, figs. 76-80. 
1934. Representative of the A. 
niger group. In the Biourge 
Collection 1939 234 

A. cimmerius Berkeley and Curtis, 
in Grevillea 3: 108. 1875. This 
specimen was identified by Dr. 
Farlow (Bibliographical Index 
I. pt. 1, page 277) as Periconia 
chlorocephala, Fres. 

A. cinerescens Bainier and Sartory, 
in Bui. Soc. Mycol. France 27: 
98-104, pi. Ill, figs. 6-12. 1911. 
In the A. glaucus group. 

A. cinereus Spegazzini, in Anal. Soc. 
Cient. Argen. 10: 162-163. 1880. 



338 



A MANUAL OF THE ASPERGILLI 



Not identifiable; believed in the 
A. glaucus group. 

A. cinnamomeus Schiemann, in 
Ztschr. Induk. Abstain, u. 
Vererbungslehre, Bd. 8, Heft 
1/2: 1-35, 16 figs. 2 pi. (1 col.). 

1912 223 

Syn. A . niger mut. cinnamomeus . . . 223 

A. cinnamominus (Weiss) Dodge, in 
Dodge Med. Myc., p. 627. 

1935 200 

Syn. S. cinnamominus Weiss, 
in Ann. Parasitol. Hum. Comp. 
8: 189-193, 5 figs. 1930. See 
A . terreus 200 

A. circinatus Mangin, in Bui. Soc. 
Mycol. France 15: 223, PI. XI, 
figs. 5, 6, 7. 1899. Not recog- 
nizable — probably not an Asper- 
gillus. 

A. citricus (Wehmer) Mosseray, in 
La Cellule XLIII: 262-263, pi. 
4, fig. 105-106. 1934. Memberof 
A. niger group. In Biourge's 
Collection 235 

A. citrino-niger Mosseray, in La 
Cellule XLIII: 231-232, pi. 1, 
fig. 7. 1934. Member of A. 
niger group. In Biourge's Col- 
lection only 233 

A. citrisporus von Hohnel, in Sit- 
zungsber. Kais. Akad., Wiss. 
Wien, Math- Naturw. Kl. 
Ill, Abt. I: 987. 1902. In the 
A. tamarii group 251 

A. citromyces E. Ruppel, in J. Pharm. 
Belg. 19: 63-8. 1937. C.A. 31: 
6687. 

A. clavatus Desm., in Ann. Sci. 
Nat. Bot. Ser. II, 2: 71, pi. 2, 
fig. 4. 1834 92 

A. clavatus var. gigantea Blochwitz. 
Cited by Biourge in unpublished 
MS. 

A. clavatus Desm. mut. gigantea 
Blochwitz, in Ann. Mycol. 
XXVIII. (3/4) : 1920 is used by 
Blochwitz on several occasions.. . 97 

A. clarellus Peck, in N. Y. St. Mus. 
Rept. 34: 44, pi. 2, figs. 1-5. 
1881. 
Syn. A . clavatus Desm 98 



S. coerulea Blochwitz, listed in 
Biourge's MS. as one of the blue 
Aspergilli (A. sydowi). 

A. concentricus Castellani, in Trans. 
Int. Derm. Cong. 6: 667-671, pi. 
49, 50. 1907. 
Syn. Epidermophyton concentricum 
(Blanchard) Castellani and 
Chalmers. 

A. condylomatae Greco. A name 
attached to a pathogen without 
data for identification. 

A. conicus Blochwitz published by 
E. Dale in Ann. Mycol. 12: 38. 
1914. Described as a Penicil- 
lium in Ann. Mycol. 10: 465. 
1912. In the A. restictus 
group 140 

A. conoideus Sprengel, in Sys. Veg. 
ed. 16, V. 4: 541. 1827. This 
is cited as Byssus conoidea Mull. 
in Flora Danica, Taf . 897, fig. 2 
which is a myxomycete, hence 
this name may be dropped. 

A. cookei Sacc, in Sylloge Fung. IV: 
71. 1886. 
Syn. A. mucoroideus Cooke, in 
Grev. 12: 9. 1883. Some mem- 
ber of the A. glaucus group. 

E. coriorum Wallr., in Compendium 
Flora Germania 4: 330. 1833. 
Syn. E. repens DeBary fide Man- 
gin. Not recognizable. 

S. corolligena Massee, in Bui. Royal 
Bot. Gard. Kew. No. 1, p. 5. 
1910. Not recognizable. 

S. coronata v. Tieghem, in Bull. Soc. 
Bot. France 24: 103. 1877. 
Unidentifiable. 

S. coronella Costantin, in Muced. 
Simples p. 34, fig. 2, 1888. Uni- 
dentifiable. 

A. crocatus Berkeley and Curtis. 
Specimen only in the Curtis 
Collection. This type specimen 
has been identified as Chon- 
dromyces and is so reported in 
Farlow's Bibliographical Index 
I. pt. 1, page 277. 
A. cucurbitaceus in the Curtis Collec- 
tion (Harvard). Was identified 
by Farlow as Choanephora. 



CHECK LIST OF SPECIES AND GENERA 



339 



A. curtisii Berkeley, in Ilavenel 
Fungi Carolinense fasc. IV. 
1855. Farlow identified this 
in his Bibliographical Index as 
Rhinotrichum curtisii. 

S. cyanea Bainier, nomen nudum, in 
Biourge's list of the nidulans 
group. 

A. cyaneus (Mattlet) Dodge, in 
Dodge Med. My col. p. 636, 1935. 
Syn. S. cyaneus Mattlet, in Ann. 
Soc. Belg. Med. Trop. 6(1): 32. 
1926. In A . sydowi series 186 

A. cyanogenes Biourge, nomen 
nudum. One of the A. conicus 
strains in A . restrictus series 140 

S. dasytricha Ell. and Ev., in Jour. 
My col. 2: 104. 1886. Sacc. 
Syllog. X: 525. Not an Asper- 
gillus. 

A. delacroixii (Sacc.) Thorn and 
Church, in The Aspergilli, p. 190. 

1926 282 

Syn. S. delacroixii Sacc, in Sylloge 

10:527 282 

Syn. S. ochracea Delacroix, in Bui. 
Soc. Mycol. France 7: 109, pi. 
VII, fig. f. 1891 282 

A. delacroixii Sacc. and Sydow, in 
Sylloge Fungorum 14: 1044. 
Syn. A. olivaceus Delacr., in Bui. 
Soc. Mycol. France 13: 118. 
1897. Some unidentifiable 

member of the A. glaucus group. 

A. densus Mosseray, in La Cellule 
XLIII: 232-234, pi. 4, figs. 99- 
102. 1934. In the Biourge 
Collection 233 

A. depauperatus Petch, in Trans. 
Brit. Mycol. Soc. XVI. p. 244, 
text fig. 1931. Parasite upon 
Lepidosaphis ulmi on Hawthorn 
in England, also on Aspidiotus 
in Ceylon. As described: coni- 
diophores up to 50m by 2.5 to 3.5m, 
broadening to a fertile apex 4 to 
5/x in diameter with a few chains 
of conidia attached directly to 
the vesicular surface; conidia 2 
to 4m by 1.5 to 2.5u. Possibly 
some strain of the A. restrictus 
series not far from A. gracilis. 



A. derxii Biourge, nomen nudum, 
listed among the A . versicolor 
group in unpublished MS. and 
represented in his collection in 
1939. 

E. desmazieri Castagne, PI. Mars. 
II, Sup., p. 56. 1851. See also 
Berlese, Fungi Moricolae Ap- 
pendice p. 11. 1889. As de- 
scribed by Berlese, this was a 
perithecial form probably be- 
longing to the A. glaucus group, 
but not sufficiently described to 
identify. 

A. desseyi Speg., in Physis., Rev. 
Soc. Argentina Cien. Nat. VIII: 
115-117, 1 fig. 1925. Named 
A. dessyi Speg., in Rev. Appd. 
Mycol. 4: 542. 1925. Some 
form near A . fumigat us 151 

A. dierckxii Biourge, nomen nudum, 
on culture distributed by 
Biourge; a strain of the A. 
repens series. See Thorn and 
Raper U. S. D. A. Misc. Publ. 
426, p. 12. 1941 107 

A. diplocystis (Sartory, Sartory, 
Hufschmitt and Meyer) Dodge, 
in Dodge Med. Mycol. p. 625. 

1935 121 

Syn. E. diplocyste Sartory, 
Sartory, Hufschmitt and Meyer, 
in Compt. Rend. Soc. Biol. 104: 
881-883. 1930. The authors 
describe a mold with conidial 
head near A. nidulans and the 
ascospores of A. chevalieri 121 

E. diplocystis B. and Br., in Jour. 
Linn. Soc. (London), Bot. 14: 
55-56, Tab. 10. 1875. Neither 
description nor figure would 
identify this with any definite 
Aspergillus, although the speci- 
men may have been the perithe- 
cia of some member of the A. 
glaucus group. 

S. dipus Ferdinandsen and Winge, in 
Bot. Tids. 30: 220, fig. 6. 1910. 
Not separable from other large- 
spored forms in this section of 
the A . niger group 228 



340 



A MANUAL OF THE ASPERGILLI 



-4. disjunctus Bainier et Sartory, in 
Bui . Soc . My c . France 27 : 346-368, 
pi. X-XI. 1911. A member of 
the A. glaucus group. SeeA.echi- 
nulatux 133 

A. diver sicolor Waksman, in Soil 
Science 2: 126. 1916. The refer- 
ence to an organism by Waksman 
under this name was a typo- 
graphical error for A. versicolor. 

S. dubia (B. & Br.) Sacc, in Fungi 
italici pi. 902, and Sylloge 4: 72. 
1886. 
Syn. A. dubius Corda, in Berkeley 
and Broome, in Ann. Nat. Hist. 
2 Ser., 7: 100 (No. 520). 1851. 
Not thus far recognized. 

A. dubiosus Lindau, in Rabh. Krypt. 
Fl. 8: 151. 1907. In the .4. can- 
didus group. 

A. eburneus Biourge, nomen nudum, 
was attached to a culture re- 
ceived from Biourge and re- 
corded as NRRL 515. It is 
accepted as A. niveus 202 

A. echinosporus Sorokin. Abst. by 
Busch, in Ztschr. f. Pflanzen- 
krankheiten 3: 155. 1893. Ref. 
in Sacc. Sylloge 11: 592. 1895. 
The description suggests a Hap- 
lographium. 

A. echinulatus (Delacr.) Thorn and 
Church, in The Aspergilli , p. 107. 

1926 131 

Syn. E. echinulatum Delacr., Soc. 
My col. de France, Bui. Trimest 
9: 266, PI. XIV, fig. III. 1893. 
Syn. E. verruculosum Vuill., in Soc. 
Mycol. de France, Bui. Trimest. 
34: 83. 1918. In A. glaucus 
group 131 

A. effusus Tiraboschi, in Ann. di Bot. 
7 (fasc. 1): 16. 1908. See also 
Thorn and Church, in Am. Jour. 
Bot. 2:109-110. 1921. Describ- 
ed originally from rotten corn 
(Zea Mays); belongs to the A. 
flavus-oryzae group 267 

A. elatior Mosseray, in La Cellule 
XLIII: 253-255, pi. 3, figs. 29-32. 
1934. In the Biourge Collection. 234 



A. elegans Gasperini, in Atti. Soc. 
Toscana Sci. Nat. Pisa, Mem. 8: 
(fasc. 2): 328. 1887. In A. 
ochraceus group 281 

E. epixylon Kunze and Schm. (D. 
Schw. No. 83; Wallr. Fl. Crypt. 
No. 2078). In Compendium 
Florae Germanicae 4: 331 . 1883. 
This was evidently some member 
of the A. glaucus group. 

A. erythrocephalus Berkeley and 
Curtis, in Jour. Linn. Soc. 
[London], Bot. 10: 362. 1869. 
In A. tamarii group 257 

Inzengaea erythrospora Borzi, in 
Jahrb. Wiss. Bot. (Pringsheim) 
16: 450-463, pis. 19-20. 1884. 
Manifestly A. variecolor 163 

A. exiguus Hann. 

Syn. A. conicus fide Blochwitz, in 
Ann. Mycol. 31: 73. 1933. 

S. ferruginea Cooke, in Grevillea 
VIII. (1879), 95., in Jour. 

Quekett Mic. Club 2 Ser. 2: 139. 
1885. Not an Aspergillus. Re- 
published in Veg. Wasps and 
Plant Worms, p. 184. 1892; see 
also Petch in Trans. Brit. Myc. 
Soc. XVI, p. 72. 1931, who ex- 
amined type specimens as rust- 
colored patches on pupae; co- 
nidophores hyaline smooth, 12m 
diam., vesicle globose 50m ; pri- 
mary sterigmata 30 by 4 to 12/*, 
secondary 14-18 by 7-9m, conidia 
from 9 by 5m to 6 to 9m subglo- 
bose, brown, coarsely rough; a 
brown member of the A. niger 
group. 

A. ferrugineus Fuckel, in Fungi 
rhenani No. 157, also Symbolae 
Mycologicae in Jahr. d. Nas- 
sauischen Vereins fur Natur- 
kunde Jahrg. XXIII and XXIV: 
358. 1869-70. Not identified. 

A. ferrugineus Link, in Sp. Plant, Ed. 
4, 6: pt. 1, p. 68. 1824. Also in 
Fries. Sys. Myc. 3, p. 387. 1829. 
Both authors seem to have had 
members of the A . glaucus group. 

A.ficuum (Reich.) Hennings 225 



CHECK LIST OF SPECIES AND GENERA 



341 



Syn. Sterigmatocystis ficuum 
(Reich.) P. Henn., in Hedwigia 
34, Heft 2: 86. 1895. 
Syn. Istilago ficuum Reichardt, 
in Verhandl. K. K. Zool. Bot. 
Gesell. Wien. 17: 335. 1867. 
A culture distributed under this 
name (Thom 142) is a rapid ox- 
alic acid producing organism. 
In the A. niger group. 

A.fiemonthi Sopp, cited as a name on 
a culture by Biourge in MS. p. 
19, in the A. flavus group. 

A. fimetarius Peck, in N. Y. State 
Mus. Bot. Rept. 42, p. 128. 
1889. Probably .4.. candidus 
group. 

A. fimeti Sacc, in Michelia 2: 543. 
1882. Inadequately described. 

A. fischeri Wehmer, in Centralb. f. 
Bakt. II abt. 18(12/15): 390-392, 
figs. 5. 1907. In the A. fumi- 

gatus group 151 

Syn. A. fumigatus-ascosporic, see 
Thom and Church, in Am. Jour. 
Bot. 5: 91-92. 1918. 

A. flavaureus Biourge, nomen nu- 
dum, listed in his MS. as a mem- 
ber of the ^4. flavus-oryzae group 
with yellow-orange reverse, as 
shown by a culture in his collec- 
tion. 

A. flavescens Wreden, in Compt. 
Rend. Acad. Sci. Paris 65: 368. 
1867. Also St. Petersb. Med. 
Zeitschr. 13: 133. 1867. Iden- 
tifications have not been satis- 
factory. Possibly in A. nidu- 
lans group. 

A. flavidus Berkeley and Broome, in 
Jour. Linnean Soc. (London), 
Bot. 14: 101. 1875. Cited by 
Saccardo in Fungi of Ceylon, 
London 1871-73, No. 913 S. 
Probably a member of the A. 
flavus group but not more closely 
identifiable. 

A. flavipes (Bainier and Sartory) 
Thom and Church, in The Asper- 
gilli, p. 155. 1926 179 



Syn. S. flavipes Bainier and Sar- 
tory, in Bui. Soc. Mycol. France 
27: 90-96, pi. Ill, figs. 1-6. 
1911 179 

A. flavo-viridescens Hanzawa, in 
Journ. Coll. Agr. Tohoku Imp. 
Univ. Sapporo 4: 232-3, PI. 21, 
figs. 1-4. 1911. In A. versicolor 
group 192 

A. flavus Link, in Obs. p. 16, 1809. 
(H. F. Link Observations in 
Ordines plantarum naturales, 
Gesellschaft Naturforschender 
Freunde zu Berlin, Magazin 3, 
1809 — commonly cited Link 
Obs.) 263 

A. flavus forma Maydis Ciferri, R., 
in Ann. Mycol. 20: 46. 1922. 

A. flavus var. japonica. cited by 
Blochwitz in Ann. Mycol. 31: 74. 
1933, without description. 

A. flavus var. viridis Blochwitz, in 
Ann. Mycol. 32: 86. 1934. Is 
vaguely designated as a non- 
tropical strain apparently dupli- 
cating A. parasiticus Speare. . . 267 

A. flavus mut. rufa Blochwitz, in 
Ann. Mycol. 27(3/4) : 201 . 1929. 
A mutation which shows grada- 
tions to pure brown from the 
green form. 

A. flavus mut. fusca Blochwitz, in 
Ber. d. Bot. Ges. 48: 576. 1928. 

Eurotium Aspergillus flavus DeBary 
and Wor. Terminology em- 
ployed by DeBary and Woronin 
(Beitr. z. Morph. u. Physiol, d. 
Pilze, III Reihe, p. 380. 1870) 
to designate relationship of the 
asexual species, A. flavus, with 
species characterized by a per- 
fect stage and included in Euro- 
tium 263 

A.foetidus n. sp 219 

Syn. A. aureus Nakazawa. A 
black Aspergillus with a strong 
musty odor 219 

A. fonsecaeus n. sp. 

Syn. S. fusca Bainier, in Bui. Soc. 
Mycol. France 27: 29, pi. 1, fig. 5. 
1880. A black Aspergillus on 



3-42 



A MANUAL OF THE ASPERGILLI 



rubber, from Fonseca in Rio de 
Janeiro 227 

A. fontoynonti Gueguen, in Archives 
de Parasitologic 14: 177-192, 2 
plates, 36 figs. 1910. See also 
Compt. Rend. Soc. Biol. Paris 
66:1052. 1909. The figures and 
description place this form un- 
questionably in the A. glaucus 
group 147 

A. freisingianus Biourge, nomen 
nudum, listed with the A. clava- 
tus group as "Tres liquifiant, 
comme celle de Blochwitz." In 
the Biourge Collection 1939. 

E.fructigenum Link, in Sp. PI. Ed. 4, 
6: 80. 1824. Some unidentifi- 
able member of the A. glaucus 
group. 

S. fuliginosa Bainier, in Bui. Soc. 
Bot. France 28: 79. 1881. Some 
variety of A. niger. 

A. fuliginosus Peck, in Bui. Buffalo 
Soc. Nat. Sci. 1: 69. 1873. Also 
in Ann. Rep. N. Y. St. Mus. Nat. 
Hist. 26: 79. 1874. Some 
strain of the A. niger group. 
Mosseray applied it to a culture 
in Biourge's Collection 233 

E. fulvescens Cooke, in Grevillea 8: 
11. 1880. 
Syn. Badhamia fulvescens Cooke. 
Change of generic designation 
only, from the same material. 
Not identifiable from descrip- 
tions. 

A. fulvus Montagne, in Ann. Sci. 
Nat. Bot. Ser. 3, Vol. XII: 298. 
1849. 
Syn.<S./uZraSacc.Syll.4:74. Not 
identifiable. Described from 
diseased silk worms in Southern 
France, but not since identified. 

A. fumaricus Wehmer, in Ber. Deut. 
Chem. Gesell. 51(14): 1663-68, 
figs. 1-6. 1918. A unique mem- 
ber of the A . niger group 227 

Sartorya fumigata Vuillemin, in 
Compt. Rend. Acad. Sci. (Paris) 
184: 136-137. 1927. Possible 
synonym of A. fischeri q.v 153 



A. fumigatoides Bainier and Sartory, 
in Bull. Soc. Mycol. France 25: 
112, pi. 5. 1909. Compt. Rend. 
Soc. Biol. [Paris] 66: 22. 1909. 
See also Sartory, A., Champig- 
nons parasites de l'homme et des 
animaux, p. 573. 1922 153 

Syn. E. fumigatoides (B. & S.) 
Sacc.SylI.22:1255. 1913. Prob- 
ably close to A. fischeri 153 

A. fumigatus Fresenius, in Beitrage 
zur Mykologie, p. 81, pi. 10, figs. 
1-11 Frankfurt. 1850-53 148 

var. Alpha, Sion and Alexan- 
drescu, in Compt. Rend. Soc. 
Biol. 64: 288-289. 1908 151 

var. cellulosae Baumli, cited by 
Sartory, Sartory, and Meyer, in 
Compt. Rend. 203: 1289-91. 
1936. See C. A. 31: 2249. 1937. 

Mut. helrola E. Yuill, in Jour. Bot. 
(London) 1939. p. 174. A 
buff-colored mutant which ap- 
peared in culture 150 

var. magnus Nakazawa, Simo, and 
Watanabe, in Jour. Agr. Chem. 
Soc. Japan No. 144. 1936. (In 
Japanese.) 

var. minimus Surtory, in Bui. 
Acad. Med. Paris Ser. 3, 82: 304. 
1919 151 

var. tumescens Blumentritt, in Ber. 
Deut. Bot. Gesell. 23: 419-427, 
PI. 19, fig. 5, 6, 18-21. 1905.... 151 

var. subglobosus Blochwitz, in Ann. 
Mycol. 33: 244. 1935. 
Aspergillopsis fumosus Sopp, in 
Videnskapselskapets Sks. I 
Mat.-Naturv. Kl. No. 11, pp. 
202-204, Taf. XX, fig. 149. 1912. 
Not identified but suggested as 
some strain near A. ustus. 
A. fungoides Greco, in Origine des 
tumeurs et mycoses Argentines, 
Buenos Aires pp. 505-509, figs. 
297, 298, 299. 1916. The de- 
scription is inadequate for iden- 
tification. 
S. fusca Bainier, in Bull. Soc. Bot. 
France 27: 29, pi. I, fig. 5. 1880. 
Compare Roumeguere Fungi 



CHECK LIST OF SPECIES AND GENERA 



343 



gallici exsiccati, No. 995 grown 
upon moist bread in Toulouse, 
1880. Some member of the A. 
niger group 227 

A. fusco-cinereus Ellis and Morgan, 
M.S. name attached to Morgan's 
specimen No. 674. Evidently a 
member of the .4. clavatus group 
but not since seen 98 

E.fuscum Pruss, in Linnaea 25: 78. 
1852. Some unidentifiable mem- 
ber of the .4. glaucus group. 

A. fuscus Anions, in Archief voor de 
Suikerindustriein Nederlandsch- 
Indie 29, Deel 1, January-June 
pp. 8-10. 1921. Not A. fuscus 
of Bonorden, Bainier, or Schie- 
mann. See: A. terreus group.. 200 

A. fuscus Bonorden, in Bot. Ztg. 
Jahrg. 19:202. 1861. Not iden- 
tifiable 224 

A. fuscus Schiemann, in Ztschr. 
Induk. Abstam. u. Vererbungs- 
lehre. Bd. 8(1/2): 1-35, 16 figs., 

2 pi. (1 col.). 1912 224 

Syn. A. schiemanni (Schiemann) 
Thorn q.v 224 

A. galactus Link, cited by Yonemoto 
and Kato, in Bull. Miyazaki 
Coll. Agr. Forestry 3: 59-66. 
1931. Perithecia formed be- 
tween 15° and 28°C. 

A. galeritus Blochwitz, in Ann. My- 
col. 27(3/4): 205, Taf. III. 1929. 
Syn. of A. terreus Thom 197 

A. gallomyces Calmette, Alb. Kaiser- 
liches Patent Amt. Klasse 12 q. 
N. 129164 August 12 (1900). Ap- 
parently a variant of A. niger. 

A giganteus (Mattlet) Dodge, in 

Dodge Med. Myc, p. 629. 1935. 225 
Syn. S. gigantea Mattlet, in Ann. 
Soc. Beige. Med. Trop 6: 35. 
1926. Not A. giganteus Wehmer. 
Some member of the A. niger 
group. 

A. giganteus Wehmer, in Centralb. f." 
Bakt. 2 abt., 18(13/15): 385. 
1907. Seethe A. clavatus group. 95 

A. giganto-sulphureus Saito, in Jour. 
Coll. Sci. Imp. Univ. Tokyo 18: 



48, pi. Ill, fig. 12a to d. 1904. 
Apparently a non-ascosporic 
member of the A. glaucus group. 
A. gigas Speg., in Ann. Mus. Nac. 
Buenos Aires Ser. 3, V. 13: 434. 
1911. Probably one of the A. 

tamarii series 256 

A. glaber Dale, nomen nudum at- 
tached by Biourge to a culture 
belonging with A. conicus. 
S. glauca Bainier, in Bui. Soc. Bot. 
France 27: 29, PI. I, fig. 3. 1880. 
Also Bui. Soc. Bot. France 28: 
77. 1881. Probably belongs in 

A. versicolor group 193 

A. glaucoides Spring, in Bull. Acad. 
Sci. Belg. 19: 560-572. 1852. 

Probably A. fumigatus 151 

A. glaucus Link, in Obs. p. 16. 1809. 

See A. glaucus group 100 

mut. alba Blochwitz, in Deut. Bot. 
Gesellsch. Ber. 50: 248-256. 
1932. See: A. niveo-glaucus 
Thom and Raper 135 

var. albida Speg., in An. Mus. Nac. 
Buenos Aires T. VI: 332. 1899.135 

var. minimus Hanzawa, in Jour. 
Coll. Agr. Tohoku Imp. Univ. 
Sapporo 4: 220. 1911. 

var. oblongispurus E. & W. Field 
Mus. Bot. 1: 88. 1896. 

var. olivascens Saccardo, in Miche- 
lia 2: 543. 1878. Distributed 
by Sacc. as No. 1376 Myco. ital. 

var. subolivaceus Ferraris, in Fl. 
ital. Crypt. P. I, fasc. 13, p. 911. 
1914. 

var. repens Corda, in Icones 5: 53. 
1842. 

Syn. A. repens DeBary q.v 103 

A. globosus Jensen, in Cornell Agr. 
Exp. Sta. Bui. 315, p. 482. 1912. 
The strain studied by Jensen 
(No. 2705) was clearly A. versi- 
color 193 

A. globosus Link 

Syn. Sporodinia aspergillus Schro- 

ter, in Sacc. Syll. 7: 207. 

A. godfrini Sartory and Roederer, 

in Assoc'n. Frangaise pour 

l'avancement des Sciences, 42nd 



344 



A MANUAL OF THE ASPERGILLI 



Session, Tunis, pp. 601-603. 
1913. Some one of the A. glau- 
cus group 133 

A. gracilis Bainier, in Bui. Soc. My- 
col. France 23: 92, PI. IX, figs. 
11-14. 1907. In A. restrictus 

series 138 

var. exiguus Bainier & Sartory, in 
Bull. Soc. Mycol. France 28: 47, 
pi. 2. 1912. According to the 
description this variety differs in 
physiological characters slightly 
from A. gracilis Bainier 140 

A. granulatus Mosseray, in La Cel- 
lule XLIII: 249-50, pi. 3, figs. 25- 
28. 1934. A member of the A. 
niger group; in the Biourge Col- 
lection 234 

A. granulosus Raper and Thorn, in 
Mycologia 36: 565-568, fig. 4. 
1944. In the A. ustus group. . . . 175 

A. gratioti Sartory, in Compt. Rend. 
Acad. Sci. (Paris) 170: 523-524. 
1920. Also in Champignons 
parasites de l'homme et des 
animaux, pp. 578-579. 1922. 
Probably some member of the A. 
fumigatus group 154 

A. Greconis Dodge, in Dodge Med. 
Myc, p. 634. 1935 
Syn. S. aurea Greco, q.v. 

A. griseus Link, in Sp. Plant. Ed. 4, 
6(1): 69. 1824. Incorrectly cited 
by Bonorden and Wehmer as 
Link Obs. 1: 69. 1809. Name 
also used by Fries and by Bonor- 
den. Not identified. 

A. guegueni was figured inBiourge's 
monograph of the Penicillia (La 
Cellule t. 33, fasc. 1, pp. 7-330. 
1923) as P. guegueni Plate XX. 
He afterward distributed it as A. 
guegueni. In A. restrictus series 139 

A. guttifer Mosseray, in La Cellule 
XLIII: 235-236, pi. Ill, fig. 53- 
57. 1934. In the Biourge Col- 
lection 233 

A. gymnosardae Yukawa, in Jour. 
Coll. Agr. Tokyo I: 362, PI. 18, 
figs. 1-7. 1911. This fungus 
was found by Yukawa under the 



name "awokabi" and is de- 
scribed by him as essential to the 
ripening of the tunafish prepara- 
tion, "katsuobushi." The di- 
mensions given are intermediate 
between those of A . flavus and A . 
oryzae, and closely approximate 
those of A. pseudo-flavus. Al- 
though we have cultures related 
to these forms, we have not been 
able to identify these intermedi- 
ates except as members of the A. 
flavus-oryzae group 266 

A. hageni Hallier, in Cattaneo Mico. 
Corp. Urn. p. 123, PI. 6, fig. 8. 

Florentin 1892 146 

Syn. Otomyces Hageni Hallier, in 
Zeitschr. Parasit. 1: 195. 1869. 
2: 22, 233, and 259, PI. 5. 1870. 
Not identifiable. 

A. halophilus Sartory, Sartory, and 
Meyer, in Ann. Mycol. XXVIII: 
362-363, PI. Ill, figs. 11-14. 
1930. Probably some nonasco- 
sporic member of the A. glaucus 
group 118 

A. helicophorus nomen nudum at- 
tached by Thaxter to a culture 
that was found to belong with A. 
ustus 174 

S. helva Bainier, in Bui. Soc. Bot. 
France 28: 78. 1881. In A. 
ochraceus group 281 

A. hennebergi Blochwitz, in Ann. 
Mycol. 33: 238. 1935. Regarded 
by Neill as one of the A. ochra- 
ceus group. See A. wentii group. 249 

A. herbariorum, species name seems 
to appear first as Mucor herbario- 
rum in Primitiae Florae Holsa- 
ticae by Fredericus Henricus 
Wiggers, 1780, republished in 
Facsimile edition No. 23 by W. 
Junk. 1925. See A. glaucus 
group 100 

Eurotium herbariorum ser. minor 
Mangin, in Ann. des Sci. Nat., 
Bot. (Ser. 9) 10: 365. 1909. See 
A . mangini 127 

Eurotium herbariorum ser. major 
Mangin, in Ann. Sci. Nat. Bot. 



CHECK LIST OF SPECIES AND GENERA 



345 



(Ser. 9) 10: 365. 1909. A.glau- 
cus group characterized by large 
ascospores. Would fall within 

A. echinulatus q.v 131 

A. heterocephalus Spring, in Bull. 
Acad. Sci. Belg. 19: 568. 1852. 
This name was given to colonies 
in a hen's egg which showed 
small heads globose and large 
heads columnar. Since no ade- 
quate figure or description was 
offered, it may be discarded as a 
nomen nudum. 
Physomyces heterosporus Harz. See 

A. atro-ruber. 
P. (Micro-aspergillus) Hickeyi, fig- 
ured by Biourge in La Cellule 
XXXIII, fasc. 1: 1-331. 1923 
(Penicillium Monograph) , proved 
to be indistinguishable from 

A . gracilis q.v 139 

A. hispidulus Sprengel, in Sys. Veg. 
Ed. 16, 4: 541. 1827. There is 
no suggestion of an Aspergillus 
in the description given. 
A. holmiensis Biourge, nomen nu- 
dum, listed among the A. versi- 
color group in Manuscript and in 
culture in his collection in 1939. 
A. hortai (Langeron) Dodge, in 
Dodge Med. My col., p. 628. 

1935 204 

Syn. S. hortai Langeron, in Bui. 
Soc. Path. Exotique 15: 383-384, 

figs. 1-3. 1922 204 

Syn. A. terreus Thorn 195 

A. humicola Chaudhuri and Sachar, 
in Ann. Mycol. 32: 97. 1934. 
Cited by Neill as A. versicolor. . 193 
A. humus Abbott, in la. St. Coll. J. 
Sci. 1: 15-36. Fig. 2a-e. 1926. 
See also Louisiana Station Bui. 
194. One of the A. ustus series. 174 
A. hypojanthinus Biourge, was pub- 
lished as Penicillium hypojanthi- 
num in Biourge Monogr. La Cel- 
lule 33: fasc. 1, pi 321-2, PI. 
XXII, fig. 130. 1923. After- 
ward he transferred it to Asper- 
gillus. A. restrictus series... 139 



A. incrassatus Spring, in Bull. Acad. 
Roy. Belg. 19: 559, fig. 2. 1852. 
Not identifiable. 

E. insigne Winter (exsiccati in Rabh. 
Fungi No. 1732), in Hedwigia 
1873 and Rabh. Krypt. Fl. 
Aufl. 1, Abt. 2, 2: 61. 1887. 
Probably not an Aspergillus. 

S. insueta Bainier, in Bull. Soc. My- 
col. France 24: 85-87, Tab. VIII, 
figs. 1-13. 1908. A. ustus series. 174 

A. intermedia Speg., in Myc. Arg. V. 
in An. Mus. Nac. Buenos Aires 
Ser. 3, T. 13: 435. 1911. In the 
A. niger group. 

A. itaconicus Kinoshita, in Botan. 
Mag. Tokyo 45: 60-61. 1931. 
Species diagnosis and figure re- 
printed in Acta Phytochim. 
(Tokyo) 5(3): 271-287. 1931. 
See A. glaucus group 142 

S. italica Sacc, in F. italici No. 109. 
1881. Also in Michelia 1: 91. 
1877. 
Syn. A. sterigmatophorus Sacc, in 
Atti. d. Soc. Ven.-Tren. d. Sc. 
Nat .'2, fasc. 2: 232, Table XVII, 
fig. 5-8. 1873. Some member 
of the A. candidus group 211 

A. janus n. sp. Raper and Thorn, in 
Mycologia 36: 556-561, fig. 1. 

1944 187 

var. brevis Raper & Thorn, in My- 
cologia 36: 561-563, fig. 2. 1944. 190 

A. japonicus Saito, in Bot. Mag. 
Tokyo 20: 61-63. 1906. In A. 

niger group 231 

var. capillatus Nakazawa, Takeda, 
and Suematu. Culture avail- 
able in Centraalbureau. 1939. 
var. or mut. grisea Blochwitz, in 
Ann. Mycol. 33: 240. 1935. It 
was a change of name only, A. 
malvaceus Mosseray. 

A.javanicus cited by Takahashi, and 
Sakaguchi, in Jour. Agr. Chem. 
Soc. Japan 1: No. 10, 1925, only 
in parenthesis after A . fumaricus 
Wehmer and apparently deemed 
asynonyn. In A. niger group. 



346 



A MANUAL OF THE ASPERGILLI 



A. jeanselmei Ota, in Ann. de Para- 
sitologic 1(2): 137-146. 1923. 
A . flavus group 269 

A. keratitis Ball, in Amer. Med. 2: 
31. 1901. This organism was 
found in an ulcer in the human 
cornea. Not identifiable 147 

A. koningi Oudemans, in Arch. 
Neerl. Ser. II, V. 7: 284, Tab. 
XIV. 1902. Not since identi- 
fied. 

A. laneus Link Obs. p. 16. 1809. 
Syn. Botrytis lanea (Bonorden) 
Sacc, in Syll. 4: 74. 

A. la?ieus in Schweinitz in Syn. Am. 
Bor. is syn. for Rhinotrichum 
curtissii Berk, in Grevillea 3: 
108. 1875. 

A. laokiashanensis Shih, in Lingnan 
Sci. Jour. 15(3): 368. 1933. 
Near A. unguis in the A. nidu- 
lans group 169 

E. lateritium Montagne, in Century 
VI. No. 35, Ann. Sci. Nat. Bot. 
3 Ser. XI: 154. 1849. Sylloge 
p. 257. One of the A. glaucus 
group with ascospores 7 to 10m 
in diameter. 

A. Lepidophyton Wehmer. 

Syn. Epidermophyton concentricum 
fide Dodge, Med. Myc. 490. 
1935. 

A . lignier'esi Cost . and Lucet , in Ann. 
Sci. Nat. IX. Bot. 2: 137, PI. 5, 
figs. 19-23. 1905. This culture 
from the lung of a penguin differs 
in cultural details from typical 
A. fumigatus especially by the 
presence of swollen groups of 
cells in the mycelium 159 

A. longobasidia Bainier, nomen nu- 
dum; a culture so labeled (Thorn 
4640.477) was received from the 
Bainier Collection through da- 
Fonseca. This was the type of 
A. bainieri Mosseray, in Ann. 
Soc. Sc. Bruxelles 54, ser. B, p. 
72. 1934. Member of the A. 
niger group characterized by 
very long primary sterigmata. . 253 



A. lova,7iiensis Biourge, nomen nu- 
dum; culture distributed by 
Biourge and appearing in NRRL 
Collection as No. 76. See Thorn 
and Raper U. S. D. A. Misc. 
Publ. 426, p. 18. 1941 117 

A. luchue?isis Inui, in Jour. Col. Sci. 
Imp. Univ. Tokyo 15: 469, PI. 22, 

figs. 1-8. 1901 230 

var. rubeolus Shih, in Lingnan Sci. 
Jour. 15(3): 374. 1933 230 

S. lutea Bainier, in Bui. Soc. Bot. 
France 27: 27. 1880. A. flavus 
group. See also Sartory and 
Jourde, in Compt. Rend. Acad. 
Sci. (Paris) 146:548-549. 1908. . 269 

A. luteus (van Tieghem) Dodge, in 

Dodge Med. Myc, p. 625. 1935. 

Syn. S. lutea van Tieghem, in Bui. 

Soc. Bot. France 24: 103. 1877. 

A. luteo-niger (Liitz) Thorn and 
Church, in The Aspergilli, p. 166. 

1926 226 

Syn. S. luteo-nigra Lutz, in Bui. 
Soc. Bot. France 53: 48-52. 
1907. Collected by A. Chevalier 
at San Thome, Africa, in fer- 
menting seeds of Theobroma 
cacao. In the ^4. niger group . 226 

A. luteo-niger van Luijk, nomen nu- 
dum, in Biourge 's MS. p. 15. 
Probably attached to a culture 
of some black Aspergillus. 

A. luteo-virescens Blochwitz, in Ann. 
Mycol. 31(1/2): 73-83. 1932. 
In A . tamarii series 256 

A. lutescens Bainier, nomen nudum, 
described by Thorn and Church 
in The Aspergilli, p. 193, 1926. 
In A. tamarii series 251 

A. lutricolor Biourge, nomen nudum, 
listed in MS. among the A. 
ochraceus group; in the Biourge 
Collection. 

A. Macfiei Dodge, in Dodge Med. 

* Mycol. p. 269. 1935. 

Syn. Stcrigmatocystis sp. Macfie, 

in Ann. Trop. Med. Parasitol. 

15:279-281. 1921. Pathogenic? 

In the A. niger group 239 



CHECK LIST OF SPECIES AND GENERA 



347 



A. macrosporus Bonorden, in Hand- 
buch d. allg. Myk. 1851, fig. 193. 
Not identifiable. 

A. malignus Lindt, in Arch. Exp. 
Path. Pharm. 25: 256-271, figs. 
1-11. 1889. In A. fumigatus 
group 153 

A. malvaceus Mosseray, in La Cellule 
XLIII: 265-266, PI. 4, figs. 134- 
135. 1934. A member of the A. 
niger group. In the Biourge 
Collection 235 

A. mangini 

Syn. A . minor (Mangin) Thorn and 
Raper, in U. S. D. A. Misc. Publ. 
426, p. 27. 1941. 
Syn. E. herbariorum ser. minor 
Mangin, in Ann. Sci. Nat. Bot. 
(Ser. 9)10:365. 1909. 
Member of the A. glaucus group 
with ascospores of intermediate 
size 127 

A. maximus Link, in Obs., p. 16. 
1809. Not an Aspergillus. 

A. maydis Quevedo, in De Agronom- 
ica Nos. 8 and 9, Buenos Aires, 
1912. See also Sartory, in 
Champignons parasites de 
l'homme et des animaux p. 532, 
1922. A species probably be- 
longing to the A. glaucus group 
described in connection with 
disease in horses 147 

Emericella medias Chowdhury and 
Mathur, in Ann. Mycol. 36: 61- 
63. 1938. Probably A. varie- 
color q.v 163 

A. medius Meissner, in Bot. Ztg. 55: 
336-344, 354-357. 1897. See 
Thorn and Raper, U. S. D. A. 
Misc. Publ. 426, p. 33, 1941 133 

A. melleus Yukawa, in Jour. Coll. 
Agr. Tokyo 1(3): 366, Taf. 
XVII. 1911. In A. ochraceus 
group 279 

A. mencieri Sartory et Flament, in 
Compt. Rend. Soc. Biol. Paris 
83:114-115. 1920. See also Sar- 
tory et Bailly in Les Myocoses 
Pulmonaires et leur Parasites 



(Paris) pp. 180-181. 1923. A. 
glaucus group; not since reported. 146 

A. michelii Preuss, in Linnaea 25: 76. 
1852. Not recognized. 

A. microcephalus Mosseray, in La 
Cellule XLIII, p. 225-227, PI. 3, 
fig. 42-48. 1934. Member of 
the A. niger group. In the 
Biourge Collection 233 

A. microsporus Boke. The descrip- 
tion and figures given by Cat- 
taneo and Oliva in Arch. Lab. 
Bot. Critt. Garovaglio 5: 123, 
PI. 6, fig. 9, 1888, have been seen. 
Not recognizable. 

A. micro-virido-citrinus Costantin 
& Lucet, in Ann. Sci. Nat. Bot. 
IX(2): 158. 1905. In A. flavus 
group 263 

A. minimus Wehmer, in Bot. Cen- 
tralb. 80: 449-461. 1899. See 
also Wehmer 's Monogr. p. 79, 
1899-1901. Wehmer's culture 
was lost. It has not been iden- 
tified since. 

S. minor Bainier, in Bui. Soc. Bot. 
France 27: 30. 1880. Not rec- 
ognizable. Possibly A. sydowi? 

A. minor (Mangin) Thorn and Raper 
in U. S. Dept. Agr. Misc. Pub. 
426, p. 27-29. 1941. See A. 
mangini 127 

A. minutus Abbott, in Louisiana Sta. 
Bui. 194. 1926. Also in Iowa 
St. Coll. Jour. Sci. 1: 15-36, figs. 
a, b, c, and d. 1926. See A. 
ustus group 174 

A. miyakoensis Nakazawa, Simo, 
and Watanabe, in Agr. Chem. 
Soc. Japan Jour. 12(9): 963-964. 
1936. See A. niger group 220 

A. mollis Bainier and Sartory, in 
Bui. Soc. Mycol. France XXVII : 
453, PI. XVI. 1911. Not A. 
mollis Berkeley. See A. glaucus 
group 129 

A. ?nollis Berkeley, in English Flora 
V, pt. 2, p. 340. 1836. Not 
identifiable. 



348 



A MANUAL OF THE ASPERGILLI 



A. mongolicus Biourge, nomen nu- 
dum. Biourge distributed un- 
der this name a culture essen- 
tially like A. echinulatus (q.v.). 
It appears in the NRRL Collec- 
tion as No. 137. See Thorn and 
Raper U. S. D. A. Misc. Publ. 
426, p. 33. 1941 132 

A. montevidensis Talice and Mac- 
Kinnon, in Soc. Biol. [Paris] 
Compt. Rend. 108: 1007-1009. 
1931. See Thorn and Raper U. 
S. D. A. Misc. Publ. 426, p. 26. 
1941. In A. glaucus group 125 

A. mouthoni Biourge, nomen nudum, 
listed by Biourge among his Les 
"Eurotium" in unpublished 
Manuscript. Probably in cul- 
ture in his collection. 

A. mucoroides Corda, in Icones 
fungorum II: 18, fig. 76. 1837. 
Probably some member of the 
A. glaucus group — unidentifi- 
able. 

A. mucoroideus Cook, in Grevillea 
XII: 9. 1883. See A. cookei 
Sacc. 

A. mulleri Berkeley, in Jour. Linn. 
Soc. XIII: 175. 1873. Not 
identifiable. 

A. mutabilis Bainier et Sartory, 
in Bui. Soc. Mycol. France 27: 
458, pi. XVII. 1911. In A. 
glaucus group 129 

A. mycetomi Villabruzzi and Gelo- 
nesi,in Annali Med. Nav. Colon. 
33: 283-308, 8 figs. 1927. 
Syn. Madurella sp.; undeter- 
minable from the data avail- 
able. 

A. nujcobanche Link, in Sp. Plant 
Ed. 4, 6: 65. 1824. A fungus 
from rotting Peziza — not identi- 
fiable. 

A. nantae Pinoy, in Compt. Rend. 
Soc. Biol. 97(19): 67-68. 1927. 
This was probably A. unguis in 
the A. nidulans group. Listed 
by Biourge as in his collection, 
1939 170 



A. nanus Montagne, in Sylloge 
Generum Specierumque Crypt, 
p. 300, No. 1112. Paris. 1856. 
Also in Saccardo Sylloge 
Fungorum 4: p. 71. Patavii 

1886. In the A . niger group 231 

A. nanus Oudemans, in Nederl. 
Kruidk. Arch. Ser. 3, 2: 1121. 
1904. Not A. nanus Mont. q.v. 
Probably some conidial form in 
the A. glaucus group. 
E. nebulosum Fries, in Sys. Myc. 3: 
334. 1832. No data are given 
to link this with Aspergillus. 
A. nicollei Pinoy, cited by Biourge 
in 1939 manuscript. He has 
apparently raised to species 
rank S. nidulans var. nicollei 

Pinoy, q.v 170 

A. nidulans (Eidam) Wint., in Rab. 

Krypt.-F1.12:62. 1884 156 

Syn. S. nidulans Eidam, in Cohn, 
Beitr. Biol. Pfeanzen 3: 392-411, 
PI. 20-22. 1883. 
forme Cesarii Pinoy, in Bul. Soc. 

Path.Exot.8:ll. 1915 170 

mut. coerulea Blochwitz, nomen 
nudum upon a culture in the- 
Biourge Collection, 1939. 
Listed in Biourge MS. 
mut. alba E. Yuill, in Jour. Bot. 
(London) 1939, p. 175, pi. 

618 159 

var. latus Thorn and Raper, in 
Mycologia 31(6): 657-9. 

1939 159 

var. Nicollei Pinoy, in Compt. 
Rend. Acad. Sci. Paris 144: 
396. 1907. This variety was 
found fruiting within human 
tissue in a subject affected with 
"Madura-foot." Listed in 
Biourge's Collection, 1939, as 

S. nicollei Pinoy 170 

Syn. S. nidulans var. Nicollei 
Pinoy, in Archives d. Para- 
sitologic 10: 437-458, PI. XL 
1906. 
Diplostephanus nidulayis (Eidam) 
Langeron, in Compt. Rend. Soc. 
Biol. Paris 87: 343-345. 1922. 



CHECK LIST OF SPECIES AND GENERA 



349 



Syn. for A. nidulans Eidam 

q.v 156 

A. niger van Tieghem, in Ann. Sci. 
Nat. Bot. Ser. 5, V. 8(4): 240. 
1867. See also Thorn and Cur- 
rie, in Jour. Agr. Res. 7: 1-15. 
1916 216 

Syn. S. nigra van Tieghem, in Bui. 
Soc. Bot. France 24: 102-103. 
1877. 

Syn. Aspergillopsis nigra (v. Tieg- 
hem) Speg., in Myc. Arg. V, in 
Ann. Mus. Xac. Buenos Aires 
(ser. 3) 13: 435. 1911. 

mut. cinnamomeus n. comb. See: 
A . cinnamomeus Schiemann 223 

var. laevis Blochwitz, in Ann. 
Mycol. 33: 249. 1935. Pro- 
posed for smooth-spored strains. 
A. niger var . fermentarius Nakazawa, 
Simo, and Watanabe, in Jour. 
Agr. Chem. Soc. Japan 144: 171- 
2 and 184. 1936 (in Japanese) ... 220 

mut. fusca Blochwitz, in Ann. 
Mycol. 32: 87. 1934. In the 
A. niger group. Such a separa- 
tion based upon conidial color is 
of doubtful validity. 

mut. Schiemanni n. comb 224 

See: A. schiemanni (Schiemann) 
Thorn. 

forma Tuebingen Schober, in the 
Centraalbureau, is apparently 
the strain used in the researches 
of Schober. 
A. niger citricus Wehmer, name on a 
culture received from Xeuberg 
(C. T. Xo. 4668.4) but without 
description. 

Syn. A . citricus Mosseray q.v 235 

S. nigra Bainier. Syn. for S. phoeni- 
cis (Corda) Patouillard and 
Delacroix, in the A. niger series. 
A. nigrescens Robin, in Histoire 
Xaturelle des Vegetaux Para- 
sites (Paris) p. 518, pi. 5, fig. 2. 
1853. Xo success has been 
made in interpreting Robin's 
organism 151 



A. nigricans Wreden, in Compt. 
Rend. Acad. Sci (Paris) 65: 368. 
1867. Probably A. niger group. 

A. nigriceps B. and C, in the Curtis 
Collection. Specimen collected 
by Charles Wright (No. 927) in 
Cuba. Cited by Cooke, M. C. 
in Grevillia 17: 21, Sept. 1888. 
A slide from this material 
prepared by Bullard and pre- 
served in the Harvard collection 
shows a characteristic organism 
of the A. niger series, not sepa- 
rable from A . niger van Tieghem. 

A. niveocandidus Lindau, in Rabh. 
Krypt. Fl. 8: 151. 1907. In the 
A. candidus group. 

A. niveo-glaucus Thorn and Raper, 
in U. S. Dept. Agr. Misc. Pub. 
426, p. 35-36. 1941 135 

A. niveus Blochwitz, in Ann. Mycol. 
27(3/4): 205-206, Taf. Ill, fig. 
2. 1929. A member of the A. 

terreus group 202 

var. major Blochwitz, in Ann. 
Mycol. 32(1/2): 86. 1934. Ap- 
parently belongs in the A. 

candidus group 211 

var. nubila Blochwitz, in Ann. 
Mycol. 32(1/2): 85. 1934. See 
A. carneus 202 

A. Nblting Hallier, in Zeitschr. 
Parasit. (Xot found) cited 
by Cattaneo and Oliva in Arch. 
Lab. Bot. Critt. Garovaglio 5: 
122. 1888. Xot recognizable. 

A. novus, nomen nudum attributed 
to Wehmer; culture in the Cen- 
traalbureau at Baarn was identi- 
fied by Thorn and Raper as A. 
pseudo-glaucus Ill 

E. obliteratum Schw. Syn. fungorum 
in Amer. Bor. X. 2725. Some 
unidentifiable member of A. 
glaucus group. 

A. oblongisporus E. and E., Xo. 760 
in Xuttall's Flora of Fayette 
County, West Virginia. Listed 
apparently by Millspaugh in 
"The Living Flora of West Vir- 
ginia" in W. Va. Geol. Survey 



350 



A MANUAL OF THE ASPERGILLI 



5(A): 32. 1913. as .4. glaucus 
var. oblongisporus. E. & W. 
Examination of this material 
shows it to be a mixture of A. 
flavus and A. repens. 

S. ochracea Bainier, in Bui. Soc. 
Bot. France 28: 78. 1881. 
Some member of the A . ochraceus 
group. 

S. ochracea Delacroix, in Bui. Soc. 
Mycol. France 7: 109, PI. VII, 

fig. f. 1891 282 

Syn. S. delacroixii Sacc, in Syi- 
loge 10: 527. Delacroix de- 
scribed his organism without 
recognizing the previous use 
of the specific name 282 

S. ochracea (Wilhelm) Schroter. 
See: A. ochraceus Wilhelm. 

A. ochraceo -ruber Sacc, in Miche- 
lia I: 77. 1877. (Saccardo, P. 
A. No. 1063 in Myco. Veneta, on 
bark of Walnut, 1876, see fig. 17 
in Fungi italici). Some conidial 
form of the A. glaucus group. 

A. ochraceus Wilhelm, Inaug. Diss. 

Strassburg. p. 66. 1877 279 

var. microspora Tiraboschi, in 
Annali di Botanica 7: 14. 
1908 281 

S. ochroleuca Speg., in Anal. Mus. 
Nac. Buenos Aires, Ser. 3, t. 
13: 434. 1911. In the A. ochra- 
ceus group 276 

A. ochroleucus Haller, in Enum. 
Method. Stirp. Helvet. Indig. t. 
I: p. 6. 1742. Hist. Stirp. t. 
III. 1768. Cited by Gasperini 
as questionable syn. for A. 
elegans. 

A. okazakii Okazaki, in Centralb. f. 
Bakt. 2 abt. 42(10/14): 225. 

1914. A. candidus group 211 

This is cited in the Sylloge 22: 
1260 as >S. okazakii (Saito) Sac- 
cardo. 

A. oligosporus Corda, in Icones III, 
Tab. II. Not an Aspergillus. 

S. olivacea van Tieghem, in Bull. 
Soc. Bot, France 24: 103. 1877. 
Not identified with A. olivaceus 



Preuss, 1852. Not identifiable. 
A. olivaceo-fuscus Mosseray, in La 
Cellule XLIII: 258-259. pi. 3, 
fig. 49-52. 1934. A member of 
the A. niger group in the 

Biourge Collection 234 

Cladosarum olivaceum Yuill and 
Yuill, in Trans. Brit, Myc. Soc. 

22: 194-200, PI. 11-13. 1938 73 

A. olivaceus Delacroix, in Bui. Soc. 
Mycol. France 13: 118-120, text 
fig. 1897 
Syn. .4. delacroixii Sacc. and 
Sydow q.v. Not A. olivaceus 
Preuss. 1852. Probably in A. 
glaucus group. 
A. olivaceus Preuss, in Linnaea 25: 

77. 1852. Not identifiable. 
A. Oniki on a culture from Okunuki 
in the Centraalbureau collection. 
No description found. Bloch- 
witz cites it in Ann. Mycol. 33: 
240. 1935. As A. ochraceus. 
A. oosporus Wallroth, as Flora 
Cyptogamica Germaniae No. 
1928, in Compendium Florae 
Germanicae 4: 296. 1833. 
Some one of the A. glaucus 
group. 
A. oriolus nomen nudum attributed 
to Biourge. Distributed by 
Biourge Collection and appears 
as NRRL No. 87. It was iden- 
tified as a strain of A . chevalieri 
by Thorn and Raper in U. S. D. 
A. Misc. Publ. 426, pi. 21. 1941. 120 
A. oryzae (Ahlburg) Cohn, in 
Jahresb. Schles. Ges fur Vaterl. 
Cultur 21: 226. 1883. In the 

A. flavus -oryzae group 261 

Syn. E. oryzae (Ahlb.) Korschelt. 
Name with incomplete descrip- 
tion of sake organism published 
in Dingler's Polytech. Jour. 

230:330. 1878 261 

A. oryzae var. basidiferens Constan- 
ts and Lucet, in Ann. Sci. Nat, 
Bot. IX (2): 167. 1905. In 
the A. flavus-oryzae group. 
Syn. A. oryzae var. basidifer Sacc. 
Syll. 22. 



CHECK LIST OF SPECIES AND GENERA 



351 



A. ostianus Wehmer, in Bot. 
Centralb. 80: 449 461. 1899. 
Monogr. pp. 117-119, Taf. II. 
No. I. 1899-1901. In the A. 
ochraceus group 283 

A. oval is perm us Link, in Obs. II, p. 
37, 1816; also in Sp. Plant Ed. IV, 
Vol. VI(1): 66. 1824. Cited as 
synonym of A. oosporus Wallr. 
(1833) without evidence of iden- 
tity or reason for redescription. 

A. panamensis Raper and Thorn, in 
Mycologia 36: 568-572, fig. 5. 
1944 242 

A. parasiticus Speare, in Hawaiian 
Sugar Planters Exp. Sta., Path. 
& Physiol. Ser. Bui. 12: p. 38, 
PI. 3 and 4. 1912. In the A. 

flavus group 266 

Syn. A. flavus var. viridis Bloch- 
witz, q.v 267 

A. penicillatus Greville, in Scottish 
Cryptogamic Flora 1: 32, plate 
32. 1823. 
Syn. A. glaucus (conidial); not 
identifiable to species. 

A. penicillatus Link, in Sp. Plant 
Ed. 4, 6(1): 69. 1824. The 
description given by Link is not 
sufficient to identify any form, 
but since he probably intended 
to cover the organism of Gre- 
ville, it may be assumed to be 
conidial A . glaucus. Sturm cites 
it as Briarea elegans without 
specifying his reasons. 
A. penicilloides Spegazzini, in Rev. 
Agrar. Veter. La Plata, p. 245. 
1896. In the A. restrictus 

series 142 

A. penicillopsis (Hennings) Racib. 
P. Hennings as Stilbothamnium 
penicillopsis P. Henn. & E. 
Nym. described in Fungi Mon- 
sunensis (Warburg-Monsunia, 
Bd. I: p. 37 (Leipzig). 1899. 
Exsiccati of type in Pathological 
collections U.S. Dept. Agr. Bur. 
of Plant Industry as : Raciborski 
No. 87, in Crypt. Paras. Java. 



See also Paras. Alg. u Pelz. Javas 

II: 7. 1900 282 

A. periconioides Sacc, in Ann. 
Mycol. 11: 320. 1913. No. 195 
in Sydow Fungi exotici exsic- 
cati collected by P. W. Graff on 
leaves of Carica in Luzon, 1912. 
Not identifiable. 
A. pemiciosus Inui., in Jour. Coll. 
Sci. Imp. Univ. Tokyo 15: 473. 
1901. Inui recorded a yellowish 
greenish color in the mycelium of 
this species which is regarded as 
related to A. luchuensis and A. 

niger 230 

A. pertardus is a nomen nudum on a 
culture distributed by Biourge. 
The organism was close to A. 

restrictus series 142 

A. petiolatus Haller, in Historia stir- 
pum indigenarum Helvetiae 
inchoata, etc. 1768. 
A. phaeocephalus Durieu and 
Montagne, in Fl. Alg. p. 342. 
1849. 
Syn. S. phaeocephala (Dur. et 
Mont.) Saccardo, Fungi italici 
fig. 903 and No. 1244 in Sacc. 
Myc.Veneta. 1877. One of the 
A. niger group. 
A., phoenicis (Corda) Thorn, in The 

Aspergilli,p.l75. 1926 222 

Syn. S. phoenicis (Corda) Patouill. 
and Delacr., in Bui. Soc. Mycol. 

France 7: 119, PI. 9. 1891 223 

Syn. Ustilago phoenicis Corda, in 
Icones Fungorum 4: 9, pi. 3, 
fig. 26. 1840. One of the A. 

niger group 223 

A. pictor Blanchard, cited by Castel- 
lani & Chalmers in Man. Trop. 
Med. p. 806. 1913. 
Syn. Trichophyton pictor Blan- 
chard, in Traite Path. Gen. II: 
919. 1896. The production of 
polychromatic cultures suggests 
relationship to A. versicolor. 
A. pollinis Howard, in Am. Bee 
Jour. 36: 577-578. 1896. See 
also idem. 38: 530-531. 1898. 
This was A. flavus fide Turesson 



352 



A MANUAL OF THE ASPERGILLI 



in Svensk Bot. Tidskr. 11: 30. 
1917 266 

S. polychroma Ferraris, in Fl. It. 

Crypt. Hyph. p. 640. 1906 193 

Syn. A . versicolor q.v 190 

A. polychromus De Mello, in Jour. 
Indian. Bot. 1(5): 158-161. 
1920. The data given are not 
sufficient to distinguish whether 
he had A. nidulans or A. 
sydowi. 

A. polychromus Sartory, Sartory, 
and Meyer. Cit. Syn. of A. 
versicolor fide Blochwitz in Ann. 
Mycol. 31: 73. 1933. 

A. polymorphous Moquin-Tandon, in 
Elements Bot. Med. 2 Ed., 469. 
1866. Not identifiable. 

A. pouchetii Montagne, in Ann. Sci. 
Nat. Bot. 4 Ser. 6(12): 182-183. 
1859. As described this was one 
of the mucors, noted as having 
resemblances to A. maximus 
(Sporodinia). 

A. praecox Mosseray, in La Cellule 
XLIII: 229. 1934. Cited as 
synonym of A. fuliginosus 
Peck 233 

*S. prasina Bainier, in Bull. Soc. 
Bot. France 27: 31. 1880. Not 
identifiable. 

A. profusus Hann, nomen nudum. 
Cited by Thorn and Raper in 
discussing A. scheelei Bainier 
and Sartory (Bui. Soc. Myc. 
France 28: 257. 1912) as nearly 
related to A . scheelei. See Thorn 
and Raper, U. S. D. A. Misc. 
Publ. 426, p. 13, 1941. Syn. of 
A. pseudoglaucus Ill 

A. proliferans Geo. Smith, in Brit. 
Mycol. Soc. Trans. 26(1/2): 26, 
PI. III. 1943. In the A. 
ruber section of the A. glaucus 
group 117 

A. pseudo -carbonari us (Bainier) 
Mosseray, in La Cellule XLIII: 
224-225, PI. 3, fig. 7-13. 1934. . . 233 
Syn. S. pseudo-carbonaria nomen 
nudum on culture (Thorn 4640.- 
482) from the Bainier collection. 



A. pseudo-citricus Mosseray, in La 
Cellule XLIII: 228-229, pi. 4, 
figs. 103-104. 1934. Member 
of the A. niger group; in the 

Biourge Collection 233 

A. pseudo-clavatus Purjewicz, in 
Schrift. Naturforch. Gesell. 
Kiev. 16, 2, p. 309, 1900. See: 
Saccardo Sylloge Fung. 16, p. 

1028. In A. clavat us group 98 

A. pseudo-elatior Mosseray, in La 
Cellule XLIII: 255-256, pi. 3, 
figs. 33-37. 1937. Member of 
the -4. niger group; in the 

Biourge Collection 1939 234 

A. pseudoflavus Saito, in Centralb. 
f. Bakt. 2 abt. 18(1/3): 34, figs 

15-18. 1907 266 

Syn. S. pseudoflava Sacc. Sylloge 
Fungorum 22: 1260-1266. The 
morphology given indicates that 
A. pseudoflavus is one of the 
intermediate forms which bridge 
the gap between typical A . flavus 
and A. oryzae. 
A. pseudoglaucus Blochwitz, in Ann. 
Mycol. 27: 207. 1929. Emended 
description by Thorn and Raper 
in U. S. D. A. Misc. Publ. 426, 
p. 12, 1941. A. glaucus group.. . . 110 
S. pseudo -nidulans Vuillemin, Arch. 
Parasitologie 8: 540-542. 1904. 
Vuillemin transfers the asco- 
sporic form described by Grijns 
as A. fumigatus in Centralbl. 
Bakt. II, 11: 330. 1903, to this 
specific name, emending Grijns's 
description by indicating the 
double nature of the band by 
which he separates his form from 
A. ?iidulans as described by 
Eidam. This discussion by 
Vuillemin tallies with the com- 
monest of our American soil 
forms of A. nidulans but not 
with the description by Grijns. 
A. pseudo-niger Mosseray, in La 
Cellule XLIII: 256-258, PI. 4, 
figs. 113-117. 1934. A member 
of the A. niger group; in the 
Biourge Collection. 1939 234 



CHECK LIST OF SPECIES AND GENERA 



353 



S. pseudo-nigra Costantin and Lucet, 
in Bui. Soc. Mycol. France 19: 
33-44. 1903. In the A. niger 
group. 
A. -pseudo -Schiemanni Biourge, 
nomen nudum, cited from Cen- 
traalbureau catalogue 1931, as 
an actively diastatic organism 
represented in the Biourge Col- 
lection. 

A. pulchellus (Speg.) Thorn and 
Church, in The Aspergilli, p. 181. 

1926 229 

Syn. Aspergillopsis pulchella 
Speg., in Myc. Arg. V, An. 
Mus. Nac. Buenos Aires, Ser. 3, 
T. 13: 436. 1911. In the A. 
niger group 229 

E. pulcherrimum Winter, in Rabh. 
Krypt. Fl. 2, aufl. 1 Abt. 2: 
60. 1887. Noted there as also 
in Herbarium of Winter and in 
Hansen, Fungi fimicoli Danici 
1041 of Sep. Abdr. A copro- 
philus form from the dung of 
foxes in Leipzig and dogs in 
Denmark, by Hansen; not an 
Aspergillus. 

A. pulmonum hominis Welcker. 
Discussed by von Dusch, in 
Virchow's Archiv. (N. F. 1) 11: 
561-566. 1857. Apparently 
A.fumigatus 151 

A. pulverulentus (McAlpine) Thorn, 
in Jour. Agr. Res. 7: 10-11. 

1916 223 

Syn. S. pulverulenta McAlpine, in 
Agr. Gaz. N. S. Wales 7: 302. 
1896. In the A. niger group 223 

A. pulvinatus B. and C. original 
collection as far as seen appears 
to be in the Curtis Collection 
from Society Hill, S. C. 1855; 
also one marked F. cub. — Wright 
No. 642 in the Curtis Collection; 
another series of specimens of B. 
and C. No. 1648 in Ellis and 
Everhart N. A. Fungi collected 
on dead twigs at Newfield, N.J. 
1885. Also No. 2306 in the Ellis 
Collection. Not an Aspergillus. 



*S. purpurea van Tieghem in Bui. 
Soc. Bot. France 24: 101-103. 
1877. Possibly A. nidulans. 
A. purpureofuscus Fries, in Sys. 
Myc. 3: 388. 1829. Probably 
the same as A. purpureofuscus 
of Schweinitz. 
A. purpureofuscus Schweinitz, in 
Synopsis fungorum in America 
boreali media degentium. 
Secundum observationes. In 
Trans. Amer. Phil. Soc, N. S. 
4: 282, No. 2680. 1834. Also in 
Saccardo Sylloge Fungorum 4: 
68, Patavii. 1886. Not an 
Aspergillus. 
A. purpureus Haller. Historia 
stirpum indigenarum Helvetiae 
inchoata, etc. 1768. Not 
recognizable. 
S. pusilla Peyronel, in I genu atmos- 
pherici dei funghi con micelio. 
Thesis. Padova p. 21. 1914. 

See : the A . niveus series 203 

A. pusillus Massee, in Kew. Bull. 
Misc. Inf. 4: 158. 1914. From 
Soil, Sudan. Not identifiable. 
A. pyri English, nomen nudum, 
name published in Research 
Studies of the State College of 
of Washington VIII (3): 127. 
1940. (Doctoral Thesis: Taxo- 
nomic and pathogenicity studies 
of the fungi which cause decay 
of pears in Washington.) Sub- 
sequent work by English led to 
recognition of the name as a 

synonym of A . niger 231 

A. quadrifidus Link, in Obs. 2: 36. 
1816. Probably not an Asper- 
gillus. 
A. quadrilineatus Thorn and Raper, 
in Mycologia 31(6): 660, fig. 3D 

and4B. 1939 160 

A. quercinus (Bainier) Thorn and 
Church, in The Aspergilli, p. 

186-187. 1926 276 

Syn. S. quercina Bainier, in Bui. 
Soc. Bot. France 28: 78. 
1881 276 



354 



A MANUAL OF THE ASPERGILLI 



A. quininae Heim., in Bui. Soc. 
My c. France 9: 239. 1894. The 
culture was found upon quinine 
solution but the description 
given will not separate it from 
A. fumigatus. 

A. racemosus Persoon, in Neues 
Mag. Bot. 1: 121. 1794. Also 
in Tentamen Disp. Meth. Fung, 
p. 41. 1797. Not recognizable. 

A. ramosus Hallier, in Ztschr. f. 
Parasit. 2: 266-269, PI. 6, figs. 
1-6. 1870. The figures and 
descriptions evidently represent 
a strain of A. fumigatus 151 

A. raulini, nomen nudum, in 
Biourge's table, probably at- 
tached to a culture in his collec- 
tion. 

A. rehmii Zukal, in Oesterr. Bot. 
Zeitschr. 43: 160, PI. II, figs. 
1-10. 1893. Zukal regarded 
this form as close to S. sulphurea 
Fresenius, but his description of 
perithecia and ascospores ex- 
cludes the A. ochraceus group. 
Blochwitz evidently believed 
that Zukal had a mixed culture; 
hence, the name would be unten- 
able. Cultures belonging to the 
A. ochraceus group have been 
distributed under the name but 
without proving their authority 281 

A. repandus Bainier and Sartory, in 
Bui. Soc. Myc. France 27: 463, 
Pi. XVIII. 1911. A. glaucus 



group. 



133 



A. repens (Cda.) DeBary, in Ab- 

handl. I. Senkenberg. Naturf. 

Gesellsch. 7: 379. 1870 103 

A. repens DeBary and Woronin, in 

Beitrage zur Morphologie und 

Physiologie der Pilzen p. 379. 

1866. 
Syn. A. glaucus var. repens Corda, 

in Icones 5: 53, Taf. II, fig. 24. 

1842 103 

E. repens var. amstelodami Vuill., 

Soc. Mycol. de France, Bui. 

Trimest.36:131. 1920 122 



A. restrictus G. Smith, in Jour. 
Text. Inst. 22: T. 115, fig. 5. 

1931 141 

A. restrictus var. B., G. Smith, in 
Jour. Text Inst. 22: T. 115, figs. 
4, 6, and 8. 1931. See A. 
restrictus series in A. glaucus 

group 141 

A. roseus Batsch, in Elenchus Fung- 
orum, p. 183, No. 58, fig. 58. 
. 1783. Cited by Link, Spec. PL, 
ed. IV, t. VI, pt. 1, p. 68. 1824. 
Also by various authors for a 
rosy or flesh-colored organism, 
and by Corda as Haplotrichum 
roseutn in Pracht-flora, PI. XI. 
A. roseus Link, in Sp. Plant, ed. IV, 
t. 6, part 1, p. 68. 1824. Link 
took the name "roseus" used 
descriptively, but not nomen- 
clatorially, by Batsch (El. Fung, 
p. 183, no. 58, fig. 58. 1783) for 
a mold presumed to have been 
an Aspergillus, by later authors. 
The name appears as No. 2724 in 
the Curtis Collection (1849) for a 
member of the A. candidus 
group. Neither descriptions 
nor specimen dating back to 
these authors fix this name for 
any definite series. 
A. rubens Green, in Boston Soc. of 
Med. Sc. 1868. Not identifi- 
able. 

A. ruber (Bremer) 114 

Syn. A. ruber (Spieckermann and 
Bremer) Thorn and Church, in 

The Aspergillill2. 1926 114 

Syn. Eurotiurn rubrum Bremer, in 
Zeitschr. f. Untersuch. d. Nah- 
rung. und Genussmittel IV. 1901, 
p. 72; also in Die fettverzehr. 
Organismen in Nahr. u. Futter- 
mitteln, " Dissert. Munster. 

1902 114 

Syn. E. rubrum Spieckermann and 
Bremer, in Landw. Jahrb. 31: 

81-128. 1902 114 

A. ruber Estienne 

Syn. Physomyces heterosporus 
Harz. 



CHECK LIST OF SPECIES AND GENERA 



355 



Syn. S. rubra (Estienne) Biourge. 
Syn. Monascus pur pit re us; cer- 
tainly not an Aspergillus. 
S. rubescens nomen nudum on a cul- 
ture in the Bainier Collection 
(Thorn No. 4640.487). A. 

flavipes group 181 

A. rufescens Berlese, in Fungi Mori- 
colae Fasc. VII. No. 4, Tav. 54, 
figs. 12-17. 1889. Probably 
conidial strain of .4. glaucus 
group. 
A. rugulosus Thorn and Raper, in 
Mycologia 31(6): 661-2; fig. 3E 

and4C. 1939 160 

A. rutilans Mosseray, in La Cellule 
XLIII. 234-235, PI. 4, fig. 86- 
90. 1934. A member of the A. 
niger group; in the Biourge 

Collection 1939 233 

E. sacchari Spegazzini, in Anales del 
Museo Nacional de Buenos 
Aires, 6, Ser. 2, 3: 244. 1899. 
Some insufficiently described 
member of the A. glaucus 
group. 
A. sachari Chaudhuri and Sachar, in 
Ann. Mycol. 32: 95. 1934. 
Blochwitz in Ann. Mycol. 33: 
240, 1935, leaves this species in 

A . quercinus 278 

A. salmoneus Biourge, nomen 
nudum, cited by Henrard in La 
Cellule XLIII: fasc. 2, p. 353. 
1934, as one of the series 
"glauci." On p. 370, idem, he 
records that single ascospores re- 
quired 3 months for germination 
and that this species was found 
to be homothallic. Identifica- 
tion to species within the A. 
glaucus group is not possible 
without more information. 
Alliospora Sapucaya Pirn, in Proc. R. 
I. Acad. 1883 and in Jour. Bot. 
1883. p. 234. One of the A. 
niger group causing rot in allia- 
ceous bulbs 9 

A. sartoryi nomen nudum was 
attached by Biourge to a culture 
of an organism close to A. 



gracilis in the A. rcstrictus 

series 139 

A. sartoryi Sydow, in Ann. Mycol. 
11: 156-160, PI. VIII. 1913. 
Possibly in .4. tamarii group. 

A. scheelei Bainier and Sartory, in 
Bui. Soc. Myc. France 28: 257- 
262, PI. X. 1912. See Thorn 
and Raper in U. S. D. A. Misc. 
Publ. No. 426, p. 12. 1941. A. 
glaucus group 107 

A. scheelei Bainier and Sartory var. 
B., idem. See A. repens series 
in A. glaucus group 107 

A. schiemanni (Schiemann) Thorn, in 
Jour. Agr. Res. 7: 13. 1916. 
See also A. fuscus Schiemann; 
name changed because previ- 
ously used 224 

S. schneggiana Biourge, nomen 
nudum, in Biourge 's MS; listed 
in the A. candidus group as 
applying to a culture received 
from Schnegg. 

A. sclerotifer Mosseray, in La Cellule 
XLIII (fasc. 2): 247-248. 1934. 
A member of the A. niger 



group. 



233 



A. sclerotiorum Huber, in Phyto- 
pathology 23(3): 306-8, fig. 1. 
1933. A heavy sclerotium- 
producing member of the A. 
ochraceus group 278 

A. sejunctus Bainier et Sartory, 
in Bui. Soc. Myc. France 27: 
346-368, Pis. X-XI. 1911. An 
ascosporic form of the A . glaucus 



group. 



114 



E. semi-ituniersum Marchal, in C. R. 
Soc. Roy. Bot. Belg. 33: 128, Pi. 
II, fig. 3. 1895. Not an Asper- 
gillus. 

A. siebenmanni Costantin and Lucet, 
in Ann. Sci. Nat. Bot. 9, 2, p. 
162. 1905. This name is based 
upon Siebenmann's description 
of an organism from the human 
ear identified by Siebenmann as 
A. flarus, but regarded by the 
describers as a separate species 



356 



A MANUAL OF THE ASPERGILLI 



based upon the description 
given by Siebenmann 266 

A. simplex Persoon, in Tent. Disp. 
Meth. Fung. p. 41. 1797. 
Tradition calls it a Penicillium. 

A. soya on a culture from Okunuki in 
the Centraalbureau collection. 
No description found; cited by 
Blochwitz in Ann. Mycol. 33: 
240. 1935, as A. flavus. 

S. skottsbergii Bresadola and Vester- 
gren no. 250 in Vestergren, 
Micromycetes rariores selecti 
Rossia baltica: ins. Osilia, Kiel- 
kond in silva abiegna prope 
Kattiel in foliis vivis Aqulegiae 
vulgaris. 1899. Distributed 
without description; examined in 
collaboration with Professor 
Thaxter at the Harvard Uni- 
versity Cryptogamic Her- 
barium; did not prove to be an 
Aspergillus. 

A. spadix Amons, in Arch. v. Suiker- 
industrie in Nederlandsch Indie 
29: 12-14. 1921. This is one of 
the A . tamarii series 257 

A. sparsus Raper and Thorn, in 
Mycologia 36: 572-574, fig. 6. 
1944 283 

A. sphaerospermus Corda, in Icones 
II: 18. 1854. No description. 

A. spiralis Grove, in Journal of Bot. 
23: 164, tab. 257, fig. 5. 1885. 
A conidial organism of the A. 
glaucus group. 

A. spirius cited by Amons in Arch, 
v. d. Suikerindustrie in Neder- 
landsch-Indie 29: 14. 1921. 
Not identifiable. 

S. spuria Schroeter, in Cohn, 
Kryptogamen Flora von Schles- 
ien 3: 2 Halfte, Lief. 1, p. 218. 

1893 201 

Syn. ? S. carnea van Tieghem. 
1877. See: A. cameus (van 
Tieghem) Blochwitz 201 

A. stellatus Curzi, in Rend. Acad. 
Naz. Lincei 19: 424-428. fig. 1. 
1934. Culture in Centraal- 
bureau list 1939. 
Syn. A.variecolor 163 



E. stercoraria Hansen, in Meddelelser 
fra den Naturhist. Foren. i. 
Kjobenhavn p. 310. 1876. 
Syn. Anixiopsis stercoraria 
Hansen, in Bot. Ztg. 55: 127-131, 
Tab. II, fig. 8. 1897. 

A. stercoreus Sacc, in Michelia 1: 78. 
1877. Also Fungi italici no. 19. 
An unidentifiable member of the 
A. glaucus group. 

A. sterigmatophorus Saccardo, in Atti 
Soc. Ven.-Tren. Sci. Nat. 2, 
fasc. 2: 232, Tab. XVII, fig. 5-8. 

1873 211 

Syn. S. italica q.v 211 

A. sirychni Lindau, in Hedwigia 
Bd. 43, Heft 5: 306-307. 1904.... 223 
Syn. A. pulverulentus McAlpine, 
1896. In the A. niger group 223 

A. subfuscus Johan-Olsen, in Medde- 
lelser fra Naturh. forening i 
Kristiania. 1885. Cited also 
in O. Johan-Olsen, Viden- 
skapselkabet i Kristiania p. 21. 
1886. Some member of the A. 
niger group 231 

A. subgriseus Peck, in Bui. Torrey 
Bot. Club, 22, 5: 210. 1895. 
Syn. E. subgriseum Peck, in Rept. 
N. Y. State Mus. Bot. p. 30. 
1910. Also in N. Y. State Mus. 
Bui. 150. 1911. There is no 
way to identify Peck's species. 

A. sulphureus (Fres.) Thorn and 
Church, in The Aspergilli, p. 

185-186. 1926 275 

Syn. S. sulphurea Fresenius, in 
Beitr.Z.Mykologie,Heft3:p.83, 
Tab. XI, fig. 30-33. 1863. Also 
Sylloge 4: 73. 1886. In A. 
ochraceus group 275 

A. sydoivi (Bainier and Sartory) 
Thorn and Church, in The Asper- 
gilli, p. 147-148. 1926 184 

Syn. S. sydowi, Bainier et Sartory, 
in Ann. Mycol. 11: 25-29, PI. III. 

1903. /I . sydowi series 184 

var. achlamydosporus Nakazawa, 
Simo, and Watanabe, in Jour. 
Agr. Chem. Soc. Japan 10(2): 
178-179. 1934. The absence of 



CHECK LIST OF SPECIES AND GENERA 



357 



Hulle cells is inadequate for 
separation 186 

A. syncephalis Gueguen, in Champ. 
Parasit. Horn. Anim. p. 165, fig. 
6. 1904. Probably A. fumi- 
gatus 151 

S. szurakiana Moesz, in Bot. Kozlem. 
19: 44-66, 13 figs. 1921. One 
of the A. candidus group. 

A. tabacinus Nakazawa, Simo, and 
Watanabe, in Jour. Agr. Chem. 
Soc. Japan 10(2): 177-178. 
1934. Appears to have been a 
strain of the A. versicolor 
series 193 

A. tamarii Kita, in Centralb. f. 
Bakt. etc., 2 Abt. 37, No. 14/16, 
pp. 432-452. 1913. A tamarii 
series 254 

A. tardior Biourge, nomen nudum, 
listed among his cultures of the 
A. restrictus group (Biourge 's 
Sec. I. Microaspergilli). 

A. terreus Thom, in Turesson, Gote, 
Svensk Botanisk Tidskrift 10: 
5. 1916. Without description; 
diagnosis Thom and Church 
Amer. Jour. Bot. 5: 85-86. 1918. 

The A . terreus series 195 

var. aureus, n. var 198 

var. Boedijni (Bloch.) n. comb. 

Thom and Raper. 
Syn. A. Boedijni Blochwitz, Ann. 

Mycol. 32(1/2): 83. 1934 197 

var. floccosus Shih, in Lingnan 
Science Jour. 15: 372, pi. 16, fig. 

3. 1936 198 

var. subfloccosus Shih, in Lingnan 

Science Jour. 15: 371, pi. 16, fig. 

4. 1936. 

A. terricola Marchal, in Rev. Mycol. 
15(59): 101-103. 1893. In A. 

tamarii series 253 

var. Americana Marchal in Thom 
and Church, Am. Jour. Bot. 8: 
125. 1921. Also in Thom and 
Church The Aspergilli, p. 192. 
1926 253 

A. thoynii. This was an undescribed 
sclerotium-producing strain of 
A. flavus sent by P. W. Graff to 



Amer. Type Cult. Collec- 
tion 266 

A. tiraboschii Carbone, in Atti d. 
Inst. Bot. Univ. Pavia Ser. II 
Vol. XIV: 320. 1912. In the 
A. sydowi -versicolor group 188 

A. tokelau Wehmer, in Centralbl. 
Bakt. I, 35: 140. 1903. Cited by 
Dubreuihl in Jour. Med. Bor- 
deaux 32: 312. 1902. Wehmer's 
culture was one of the A . glaucus 
group but not the pathogenic 
organism causing the disease 
"tokelau." Dodge, Med Myc. 
490. 1935. calls it Epider- 
mophyton. 

S. tropicalis Matta, in Bol. Inst. 
Brasil. Sci. 3: 51-54, 2 figs. 
1927. Probably A. sydowi but 
not separable from other strains. 

A.tubingensis (Schober) Mosseray, in 
La Cellule XLIII: 245-247, pi. 
3, fig. 58-60. 1934. Member 
of the A. niger group; in the 
Biourge Collection 1939 234 

A. tunetanus (Langeron) Dodge, in 
Dodge Med. Mycol., p. 635. 
1935. 
Syn. S. tunetana Langeron, in Bui. 
Soc. Path. Exot. XVII: 345-347, 
illust. 1924. See A. sydowi 
series 186 

A. umbrinus Patterson, in Bull. 
Torrey Bot, Club 25: 284. 1900. 
In the A. tamarii series. 

A. umbrosus Bainier and Sartory, in 
Bui. Soc. Mycol. France 28: 267, 
PI. XII. 1912. In the A. 
glaucus group 129 

A. unguis (Emile-Weil and Gaudin) 
Emend. Thom and Raper in 
Mycologia 31(6): 667-8, fig. 6. 

1939 169 

Syn. S. unguis Emile-Weil and 
Gaudin, in Arch. Med. Exp. 
Anat. Path. (Paris) 28: 463-465. 
1919. See Thom and Raper, 
The A. nidulans group, Mycolo- 
gia 31: 667. 1939 169 

A. ustus (Bainier) Thom and Church, 
in The Aspergilli, p. 152-153. 
1926 171 



358 



A MANUAL OF THE ASPERGILLI 



Syn. S. usta Bainier, in Bui. Soc. 
Bot, France 28: 78. 1881. .4. 

iistus group 171 

var. laevis Blochwitz, in Ann. 
Mycol. 32(1/2): 84. 1934 175 

A. ustilagu Beck, in Itin. Prin. S. 
Coburgi. 2: 148 (Wien). 1888. 
Saccardo, in Sylloge Fungorum. 

10:526 (Patavii). 1892 223 

Syn. A. phoenicis in the A. niger 
group 222 

A. Vancampenhouti Mattlet, in 
Ann. Soc. Beige Med. Trop. 4: 
167-176. 1924. Name only 
ibid. 6: 31. 1926. In the A. 
versicolor group 193 

S. varia Bainier, in Bui. Soc. Bot. 
France 27: 30. 1880. Probably 
some nonascosporic A. nidulans, 
or A. unguis. 

A. variabilis Gasperini, in Atti Soc. 
Toscana Nat. Sci. Pisa, Mem. 
8, fasc. 2, p. 326. 1887. From 
the description, this was proba- 
bly some strain of the A. flavus 
group 266 

A. varians Ceni, in Ri vista speri- 
mentale di Freniatria, p. 31. 
1905. Certainly identified by 
Tiraboschi in Annali di Bot. 7: 
9-10, 1908, as A. versicolor. 

A. varians Wehmer, in Bot. Centralb. 
80: 460-1. 1899. Also in Weh- 
mer Monogr. pp. 77-79, taf. I. 
1899-1901. If Wehmer's de- 
scription is correct, his species is 
not known in culture now. The 
authors have believed that A. 
varians is identical with A. 
itaconicus 142 

A. variecolor (Berk, and Br.) Thorn 
and Raper, in Mycologia 31: 663- 

667, fig. 4D and fig. 5. 1939 163 

Syn. Emericella variecolor Berk, 
and Br., in Introd. Crypt. Bot. 
p. 340-341, fig. 76. 1857. See 
Patouillard in Bull. Soc. Myc. 
Fr. 7: 43-49, pi. 4, fig. 6-12. 

1891 163 

Syn. Inzengaea erythrospora Borzi, 
Jahrb. Wiss. Bot. (Pringsheim) 



16: 450-463, pi. 19, 20 (1884). 

1885 163 

Syn. Emericella medias Chowdhury 
& Mathur. Ann. Myc. 36: 61-63. 

1938 163 

Syn. .4. stellatus Curzi, Rend. 
Acad. Naz. Lincei 19, p. 424-428, 
fig. 1. 1934 163 

A. variegatus Mosseray, in La Cellule 
XLIII: 238-239, pi. 4, fig. 72-75. 
1934. Member of the A. niger 
group; in the Biourge Collection. 
1939 233 

A. velutinus Mosseray, in La Cellule 
XLIII: 252-253, pi. 3, fig. 38-41, 
1934. Member of the A. niger 
group; in the Biourge Collection. 
1939 234 

S. veneta Massalongo, in Bui. Soc. 
Bot. Ital. No. 7-8, p. 159. 1900. 
Not cultivated and not identifi- 
able by description, although 
Werkenthin (Phytopathology 6: 
247-249. 1916) used A. venetus 
for strains now known to be A. 
terreus. 

E. verruculosum Vuillemin, in Bui. 
Soc. Myc. France 34: 83. 1918. 
See A. echinulatus in A. glaucus 
group 131 

A. versicolor (Vuillemin) Tiraboschi, 

in Ann. Bot. (Rome) 7: 9. 1908. 190 
Syn. <S. versicolor Vuillemin, cited 
by Mirsky, in These de Med. 
Nancy No. 27, p. 16. 1903. A. 

versicolor series 190 

mut. coerulea Blochwitz, in Ann. 
Mycol. 27(3/4): 201. 1929. Rep- 
resents a pure blue strain arising 
from a green strain. Probably 
A. sydowi. 
var. fulvus Nakazawa, Takeda and 
Suematu, culture available from 
the Centraalbureau, 1939. 
var. glauca Blochwitz, in Ann. 
Mycol. 32: 86. 1934. A strain 
greener than typical for the 
species, from human skin accom- 
panying a Trichophyton. Simi- 
lar green strains have been 
observed by us 192 



CHECK LIST OF SPECIES AND GENERA 



359 



A. violaceo, cited by Biourge in his 
MS. p. 16, but no data offered. 
.4. violaceo-fuscens (Was this A. 
violaceo-fuscus?) mut. grisea 
Blochwitz, in Ann. Mycol. 32: 
87. 1934. The grounds of sepa- 
ration are vague. 
A. violaceo-fuscus Gasperini, in Atti 
Soc. Toscana Sci. Nat. Pisa, 
Mem. 8, fasc. 2, p. 326. 1887. 

In .4. niger group 231 

A. virens Link, in Obs. Ord. PI. Nat. 
p. 16, 1809; also Sp. PL, Ed. 4, 6: 
pt. 1, p. 67. 1824. This has 
never been surely identified. 
A. virens (Link?) rLichelbaum, in 
Verh. Naturw. Ver. Hamburg 3 
Folge. XIV. p. 34. 1906. This 
might have been close to A. 
fischeri. 
Syn. E. virens (Eichelb.) Sacc. 
Saccardo cites description of 
preceding under this name in 
Sylloge Fung. 22: 1255. 1913. 
A. riridans nomen nudum attached 
by Biourge to a culture distrib- 
uted by him. One of the A. 

rest rictus series 1-10 

.4. riridis apparently undescribed, 
appears only on a specimen in 
the Curtis Collection now in the 
Cryptogamic Herbarium of 
Harvard University; the label is 
"A. riridis Schw. in herb., A. 
virens Lk. in Syn.: U. S., Herb. 
Schw." No Aspergillus could 
be found in the specimen. 
,4. rirido-griseus Costantin and 
Lucet, in Ann. Sci. Nat. Ser. IX, 
2: 140. 1905. One of the A. 

fumigatus group 151 

S. ritellina Ridley, in Jour. Bot. 34: 
152, Plate 357, figs. 14-16. 1896. 
No one has since reported Kid- 
ley's organism with certainty 
but a culture belonging in the A . 
ochraceus group appears under 



the name in C. B. S. list. Bloch- 
witz regarded it as a synonym for 
A. flavus 276 

.4. oulpinus nomen nudum was dis- 
tributed by Biourge. It be- 
longed to A. tamarii Kita 256 

S. welwitschiae (Bresadola) Hen- 
nings. Cited by Wehmer in 
Centralb. f. Bakt. etc., 2 Abt . 18: 
394-395. 1907. Examination of 
material from Hennings by Weh- 
mer furnished no basis for sepa- 
rating the form from A. niger. 
Examination of similar specimen 
in the Farlow Herbarium shows 
primary sterigmata up to 50 by 
12m which would place it near A. 
phoenicis. 
Syn. Ustilago welwitschiae Bresa- 
dola. 

A. wehmeri Constantin and Lucet, 
in Ann. Sci. Nat. Bot. Ser. IX, 
2: 162. 1905. The name is pro- 
posed by Costantin and Lucet 
for the organism which Brefeld 
and Wehmer described as A . fla- 
vus Link and which is so used in 
this paper. The uncertainties in 
the identification of Link's 
species do not seem important 
enough to justify the change of 
name. A. wehmeri is to be re- 
garded as a synonym of A. 

flavus q.v 266 

A. wentii Wehmer, in Centralbl. 
Bakt. 2 Abt., 2: 150. 1895. See 
also Wehmer, Die Pilzgattung 
Aspergillus, etc., in Mem. Soc. 
Phys. d'Hist. Nat. Geneva 33: 

part 2, p. 119. 1899-1901 246 

var. minimus Nakazawa, Takeda, 
Okada, and Simo, culture avail- 
able from the Centraalbureau in 

1939 249 

A. westendorpii Sacc. and March, in 
Rev. Mycol. 7: 149. 1885. In 
A. clavatus group 98 



Chapter XXV 



ACCEPTED SPECIES, VARIETIES, AND 

MUTATIONS 



A. alliaceus Thom and Church 

A. amstelodami (Mang.) Thom and 
Raper 

A. atropurpureus Zimmerman 

A. avenaceus G. Smith 

A. awamori Nakazawa 

A. butyracea Bainier 

A. caespitosus Raper and Thom. . . 

A . candidus Link 

A. carbonarius (Bain.) Thom 

A. carneus (v. Tiegh.) Bloch., emend. 

.4. carnoyi (Biourge) Thom and 
Raper 

A. chevalieri (Mang.) Thom and 
Church 

A. chevalieri (Mang.) Thom and Ch. 
var. intermedins Thom and Ra- 
per 

A. citrisporus von Hohnel 

A . clavatus Desm 

A. conicus Blochwitz 

A. delacroixii (Sacc.) Thom and 
Church 

A. echinulatus (Delacr.) Thom and 
Church 

A. effusus Tiraboschi 

A. elegans Gasperini 

A. fischeri Wehmer 

A. flavipes (Bain, and Sart.) Thom 
and Church 

A . flavus Link 

A. foetidus n. sp 

A. fonsecaeus n. sp 

A. fumaricus Wehmer 

A. fumigatus Fresenius 

A. fumigatus (Fres.) mut. helvola 
Yuill 

A. giganteus Wehmer 

A . gracilis Bainier 

A. granulosus Raper and Thom 

A. humicola Chaudhuri and Sachar. 

A. itaconicus Kinoshita 

A. jarius Raper and Thom 



244 

122 
226 
246 
220 
282 
166 
207 
229 
201 



282 

131 
267 
281 
151 

179 
263 
219 
227 
227 
148 

150 
95 
138 
175 
193 
142 
187 



A 
A 
A 
A 
A 
A 
A 
A 

A 
A 



134 A 
A 

118 

A 

121 A 

251 A 
92 

140 A 



A 
A 
A 
A 
A 
A 
A 
A 
A 
A 
A 
A 
A 

A 
A 
A 

A 
A 



janus var. brevis Raper and Thom 190 

japonicus Saito 231 

luchuensis Inui 230 

lutescens (Bain.), Thom & Church 251 

mangini n. comb 127 

medius Meissner 133 

melleus Yukawa 279 

micro-virido-citrinus Cost, and 

Lucet 263 

miyakoensis Nak., Simo & Wat.. 220 
montevidensis Talice and Mac- 
Kinnon 125 

nidulans (Eidam) Wint 156 

nidulans (Eidam) Wint. mut. alba 

Yuill 159 

nidulans (Eidam) Wint. var. 

latus Thom and Raper 159 

niger van Tieghem 216 

niger v. Tiegh. mut. cinnamomeus 

(Schiem.) n. comb 223 

niger v. Tiegh. mut. schiemanni 

(Schiem.) n. comb 224 

niveo-glaucus Thom and Raper. . 135 

niveus Bloch., emend 202 

ochraceus Wilhelm 279 

oryzae (Ahlburg) Cohn 261 

ostia7ius Wehmer 283 

panamensis Raper and Thom. . . . 242 

parasiticus Speare 266 

. penicilloides Speggazzini 142 

penicillopsis (Hennings) Racib.. 282 

phoenicis (Cda.) Thom 222 

proliferans G. Smith 117 

. pseudoglaucus Bloch 110 

, pulchellus (Speg.) Thom and 

Church 228 

. pulverulentus (McAlpine) Thom 223 
. quadrilineatus Thom and Raper 160 
. quercinus (Bain.) Thom and 

Church 276 

. repens (Cda.) DeBary 103 

. restrictus G. Smith 141 



360 



ACCEPTED SPECIES, VARIETIES, AND MUTATIONS 



361 



A. ruber (Brem.) 114 

A. rugulosus Thorn and Raper 160 

A . sclerotiorum Huber 278 

A. sparsus Raper and Thorn 283 

A. sulphureus (Fres.) Thorn and 

Church 275 

A. sydowi (Bain, and Sart.) Thorn 

and Church 184 

A. tamarii Kita 254 

A . terreus Thorn 195 

A. terreus Thorn var. aureus n. var.. 198 
A. terreus Thom var. boedijni n. var. 197 
A. terreus Thom var. floccosus Shih 198 



A . terricola Marchal 253 

A. terricola var. americana Marchal 253 
A. umbrosus Bainier and Sartory. . . 129 
A. unguis (Emile-Weil and Gaudin) 

Thom and Raper 169 

A. ustus (Bainier) Thom and Church 171 
A. ustus (Bain.) Thorn and Ch. var. 

laevis Blochwitz, n. comb 175 

A. variecolor (Berk, and Br.) Thom 

and Raper 163 

A. versicolor (Vuill.) Tiraboschi .... 190 

A . violaceo-fuscus Gasperini 231 

A , wentii Wehmer 246 



INDEX 



Abbott, E. V., 174, 175, 319 

Accepted species, 360 
Acid production 

Aconitic, 299 

Amino, 299 

Aspergillic, 272 

Citric, 237, 249, 290-293 

Fumaric, 237, 293-294 

Gallic, 237, 294 

General, 294-295 

Glaucic,299 

Gluconic, 238, 295-297 

d-Gluconic, 299 

Glucuronic, 299 

Glycolic,299 

Glyoxylic,299 

Itaconic, 143, 204-205, 297 

Kojic, 249, 258, 270, 297-298 

Malic, 299 

Melleic,299 

Oxalic, 238, 298-299 
Aconitic acid, 299 
Actinomycetes, 42, 31 
Agar slant cultures, 51 
Albino forms, 206, 212 
Alcohol, 316 

Alcoholic fermentations, 270 
Alexander, D. F., ix 
Alkaline reaction by A. clavatus, 95 
Allergy, 154, 271 
Alliospora, 9 
Alphabetical check list of species and 

genera, 330 
Amann, J., 47, 319 
Amino acid formation, 299 
Anastomoses, 65 
Anslow and Raistrick, 154, 319 
Antibiotics, 98 

from A. clavatus, 98 

from A. giganteus, 99 

from A.flavipes group, 182 



from A. flavus-oryzae group, 272 
from A.fumigatus, 154 
Arlington Farm, 227 
Ascogone, 27 
Ascomycetes, 6 
Ascophora nigrans, 9 
Ascospore, 28 

Color in A. nidulans group, 28, 156 
Germination, 28, 29 
Markings, 108, 130, 152, 162 
in A. fischeri, 162 
in A. glaucus group, 108, 130 
in A. nidulans group, 162 
Aspergillaceae, 6 
Aspergilleae, 6 
Aspergillic acid, 272 
Aspergilline, 22 
Aspergillopsis Sopp, 9 
Aspergillopsis Spegazzini, 8 
Aspergillosis, in birds, 148, 154 
Aspergillus Micheli, 6, 7 
Aspergillus, Generic diagnosis, 7 
Aspergillus species, *see also Check list 

of species, p. 331 
A. alliaceus Thorn and Church, 244*, 246, 

245, 249 
A. amstelodami (Mangin) Thorn and 

Church, 122*, 124, 106, 108, 123 
A. archiflaripes Bloch., 181 
A. atropurpureus Zimm., 226* 
A. avenaceus Geo. Smith, 246*, 243, 249 
A. awamori Nakazawa, 220* 
A. butyracea (Bainier) n. comb., 282* 
A. caespitosus Raper and Thorn, 166*- 

168, 167 
A. candidus Link, 207*-212, 208, 209 
A. capitatus ochroleucus Micheli, 3 
A. capitulo pulla Micheli, 3, 214 
A. carbonarius (Bainier) Thorn, 229*- 

230, 217, 218, 236 



* Names are cited in this index for recognized or historically important species 
only. For a complete list of published names for the Aspergilli, see the Check 
List of Species, Chapter XXIV. 

363 



364 



INDEX 



A. carneus (van Tiegh.) Bloch., 201*- 

202, 199, 200 
A. carnoyi (Biourge) Thorn and Raper, 

134*-135 
A. chevalieri (Mangin) Thorn and 

Church, 118M20, 104, 106, 108, 119 
A. chevalieri (Mangin) var. intermedins 

Thorn and Raper, 121*, 119 
A. citrisporus von Hohnel, 251*, 51 
A. clavatus Desm., 92*-95, 93, 94 
A. conicus Blochwitz, 140* 
A. delacroixii (Sacc.) Thorn and Church, 

282* 
A. echinulatus (Delacr.) Thorn and 

Church, 131*-132, 128, 130, 106, 109 
A. effusus Tiraboschi, 267*-269, 268, 69 
A. elegans Gasperini, 281* 
A.fischeri Wehmer, 151M53, 149, 152 
A. flavipes (Bainier and Sartory) Thorn 

and Church, 179 *-l 81, 180 
A.flavus Link, 263*-266, 264, 265, 69 
A.foetidus n. sp., 219*, 217, 40 
A.fonsecaeus n. sp., 227*-228, 76, 221 
A.fumaricus Wehmer, 226* 
A.fumigatus Fresenius, 68, 148M51, 149 
A.fumigatus (Fres.) var. helvola, 65, 72, 

150 
A. giganteus Wehmer, 95*-97, 96, 239 
A. glaucus Link, 3, 4, 101 
A. gracilis Bainier, 138*-139 
A. granulosus Raper and Thorn, 175*- 

178, 176, 177 
E. herbariorum, 4 

A. humicola Chaudhuri and Sachar, 193* 
A. itaconicus Kinoshita, 142*-143, 109 
A. janus Raper and Thorn, 45, 92, 187*- 

190, 188, 46 
A. janus var. brevis Raper and Thorn, 

190* 
A. japonicus Saito, 230* 
A. luchuensis Inui, 230* 
A. lutescens (Bain.) Thom and Church, 

251*-253,252 
A. mangini (Mangin) n. comb., 127*-129, 

128 
A. medius Meiss., 133*-134, 46, 128 
A. melleus Yukawa, 279* 
A. micro-virido-citrinus Cost, and Lucet, 

263* 
A. miyakoensis Nak., Simo, and Wat., 

220-221 



A. montevidensis Talice and MacKinnon, 

125*, 123 
A. nidulans (Eidam) Wint. 156*-159, 

157, 158, 161, 162 
A. nidulans mut. alba Yuill, 65, 72, 158, 

159* 
A. nidulans var. latus Thom and Raper, 

159* 
A. niger van Tieghem, 216*, 217, 218, 221, 

222, 18, 72, 239 
A. niger mut. cinnamomeus (Schiem.) n. 

comb., 73, 74, 206, 223*-224, 244, 242, 

236 
A. niger mut. schiemanni (Schiem.) n. 

comb., 73, 74, 206, 224*-225, 217, 244, 

242, 236 
A. niveo-glaucus Thom and Raper, 135*- 

137, 19, 128, 136 
A. niveus Bloch., emend. 202*-204, 199, 

200 
A. ochraceus Wilhelm, 279*-281, 280 
A. oryzae (Ahlburg) Cohn, 261*-263, 262, 

264, 69 
A. ostianus Wehmer, 283* 
A. panamensis Raper and Thom, 242*- 

244, 243, 249 
A. parasiticus Speare, 266*-267, 268, 69 
A. penicilloides Speg., 142* 
A. penicillopsis (Hennings) Racib.,282*- 

283 
A. phoenicis (Cda.) Thom, 222*, 223, 

218, 239 
A. proliferans G. Smith, 117* 
A. pseudo -glaucus Bloch., 110*— 111 , 105 
A. pulchellus (Speg.) Thom and Church, 

228* 
A. pulverulentus (McAlpine) Thom, 223* 
A. quadrilineatus Thom and Raper, 160* 

161,162 
A. quercinus (Bainier) Thom and 

Church, 276*-278, 277 
A. repens (Cda.) De Bary, 103M09, 104, 

105, 106, 108, 109 
A. restrictus G. Smith, 141*, 136, 139, 109 
A. ruber (Bremer) n. comb, 114*-117, 104, 

106, 108, 113, 115 
A. rugulosus Thom and Raper, 160M63, 

158, 161, 162 
A. sclerotiorum Huber, 278*-279, 277 
A. sparsus Raper and Thom, 283*- 
285, 284 



INDEX 



365 



A. sulphureus (Fres.) Thorn and Church, 

275*-276 
A. sydowi (Bain, and Sart.) Thorn and 

Church, 184M86, 185 
A. tamarii Kita, 254*-257 
A. terreus Thorn, 66, 195M97, 196, 199, 

200 
A. terreus var. aureus n. var., 66, 67, 

198*, 199 
A. terreus var. boedijni (Bloch) n. comb., 

66, 67, 197* 
A. terreus var. floccosus Shih, 66, 67, 198* 
A. terricola Marchal, 253* 
A. terricola var. americana Marchal, 

253*-254,252 
A. umbrosus Bain, and Sart., 129*— 131 , 

128, 130 
A. unguis (Emile-Weil and Gaudin) 

Thorn and Raper, 169M70 
A. ustus (Bainier) Thorn and Church, 

171M75, 172, 173, 176 
A. ustus var. laevis Bloch., 175*, 173 
A. rariecolor (Berk, and Br.) Thorn and 

Raper, 163M66, 158, 161, 162, 164 
A. versicolor (Vuill.) Tiraboschi, 190*- 

193, 185, 191 
A. violaceo-fuscus Gasperini, 231*, 217, 

218 
A. wentii Wehmer, 246*-247, 248, 249 
Assumptions, Basic, 10 

B 

Baarn, Holland, — See Centraalbureau 
Bacillus anlhracis, 154 
Bacteria in Aspergillus cultures, 59 
Bactericidal agents — See Antibiotics 
Bainier, G., vii, 4, 174, 227, 229, 236, 278 
Bainier and Sartory, 107, 319, 114, 129, 

133 
Barham and Smits, 270, 298 
Barnes, B., 74, 144, 145 
Barthel, C, 56 

Bary, A. De, vii, 4, 101, 26, 28, 7, 27, 103 
Basidia, 23 

Berkeley, M. J., 163, 165 
Bernhauer, K. I., 237, 293, 295, 299, 320, 

291,238,306 
Bernton,H.S.,154,320 
van Beyma, F. H., viii 



Bibliographies, types included, viii 
Check list of species with bibliographic 

references, 331-359 
General, viii, 319-330 
Topical, viii, 289-318 

Bichloride of Mercury, 61 

Binocular, wide-field, 40, 45, 51 

Biourge, Ph., ix, 4, 135, 139, 214, 232, 235 

Bisby,G.R.,ix 

"Black Aspergilli" — see A. niger group 

Blakeslee, A. F., 41, 320, 229, 230 

"Blase", 22 

Blochwitz, Adalbert, ix, 4, 64, 66, 137, 
175, 197, 202, 203, 206, 229, 231, 236, 
256, 267, 279 

Bonner, J. T., 242 

Borzi, A.,165, 163,321 

Brant, Nancy, ix 

Brefeld,0.,4 

Bulb disease, 246, 249 

Buller, A.H.R.,65,321 

Bush and Goth, 272, 300 



Calam, Oxford and Raistrick, 204, 321 
Candidus group, 206-213 

Group relationships, 212 

Occurrence, 213 

Other white Aspergilli, 206, 207, 212 

Outstanding characters, 206 

Sclerotia, 212 

Variation in head size, 209 
Capitulum, 3 
Carbon dixoide ice, 53 
Cardozo, D.M.,228 
Cathode rays, 75 
Cellulose, decomposition, 150 
Centraalbureau voor Schimmelcultures, 

viii, 231, 247, 261, 166 
Chaetomiwn, 239 
Challenger etal., 270, 197 
Characters, Diagnostic, 82 
Chemistry of Mold Tissue, 301-302 
Chitin, 317 
Chlamydospores, 28 
Chowdhuri and Mathur, 163, 321 
Christensen, L. M., 271, 304 
Chromotaxia, Saccardo, 242, 273 
Church, M. B., ix 
Ciferri,R., 166,321 



366 



INDEX 



Citric acid, 237, 249, 290-293 

from A . niger, 237. See also 290-293 

from A . wentii, 249 
Citrinin, 205 

Cladosarum olivaceum Yuills, 9, 65, 73, 
240 

Barnes "Creamy", 74, 145 

Conidium formation, 73 

Morphology of, 73 

Nuclear behavior, 73, 74 
Clarke, F. E., 198, 199 
Classification, 6 
Clavacin, 98 
Clavatin, 99 
Clavatus group, 92-99 

Antibiotics, 98-99 

Occurrence, 98 

Outstanding characters, 92 

Synonyms, 98 
Coffee fermentation, 286 
Coghill, R. D., ix, 75, 326 
Colony characters, 11-16 
Colony types, 11-13,12 
Color, 13 

in conidial walls, 14 

in conidiophores, 14, 22, 148, 153, 171, 
179, 273 

as group characters, 13 

in the mycelium, 13 

influence of pH, 14 

in the substratum, 15 
Color photographs of Aspergilli, Plates 
I-VII 

of A . alliaceus Thorn and Church, Plate 
VI, D 

of A. amstelodami (Mang.) Thorn and 
Church, Plate III, E 

of A. avenaceus Smith, Plate VI, E 

of A. candidus Link, Plate VI, A 

of A. carneus (v. Tiegh.) Blochwitz, 
Plate V, F 

of A. clavatus Desm., Plate III, A 

of A, flavipes (Bain, and Sart.) Thorn 
and Church, Plate IV, F 

of A. flavus Link, Plate VII, D 

of A. fumigatus Fres., PlatelV, B 

of A. giganteus Wehmer, Plate III, B 

of A. janus Raper and Thorn, Plate I, 
E and F; Plate V, B 

of A. nidulans (Eidam) Wint., Plate 
I,A-D;PlateIV, C 



Color photographs of Aspergilli — Cont'd. 
of A. niger group, N.R.R.L. 67, Plate 

VI, B 
of A. niger group, tan-spored mutant 

Plate VI, C 
of A. niveo-glaucus Thorn and Raper, 

Plate III, F 
of A. ochraceus Wilhelm, Plate VII, F 
of A. oryzae (Ahlb.)Cohn, Plate VII, C 
of A. quercinus (Bain.) Thorn and 

Church, Plate VII, E 
of A. repens (Cda.) De Bary, Plate III, 

C 
of A. restrictus Smith, Plate IV, A 
of A. ruber (Bremer), Plate III, D 
of A. sydoivi (Bain, and Sart.) Thorn 

and Church, Plate V, A 
of A. tamarii Kita, Plate VII, B 
of A. terricola var. americana Marchal, 

Plate VII, A 
of .4. terreus Thorn (unirradiated), 

Plate II, A; Plate V, E 
of A. terreus (U-V mutants), Plate II, 

B-F 
of A. ustus (Bain.) Thom and Church, 

Plate IV, E 
of A. variecolor (Berk, and Br.) Thom 

and Raper, Plate IV, D 
of A. versicolor (Vuill.) Tiraboschi, 

Plate V, C and D 
of A. wentii Wehmer, Plate VI, F 
Conidia, 18, 19, 24 
Coloration, 13 
Endogenous, 25 
Formation, 23, 24 
Germination, 29 
Nuclear behavior, 23 
Conidiophore, or stalk, 17, 18, 19 
Color, 22 
Surface, 19 
Connective, 25 
Contaminants, 57-62 
Bacteria, 59 

Elimination of, 41-42, 58, 59 
Mold disease, 59, GO 
Other molds, 58 
Recognition of, 58 
Cook and Lacey, 272, 301 
Corda, A.C. I., 3, 223 
Coremia, 13 
Cramer, C, 4, 239, 321 



INDEX 



367 



Culture Collections, 50 

American Type Culture Collection, 
Washington, 50 

Centraalbureau voor schimmelcul- 
tures, Baarn, Holland, 50 

National Type Culture Collection, 
London, 50 

Northern Regional Research Labora- 
tory, Peoria, 111., 50 

Thorn Collection, 50 
Culture media, See Media 
Culture, purification of, 41, 42 
Cultures, types of, 38 

Dilution cultures, 40 

Hanging drop, 45 

Single-colony inoculation, 39, 40 

Single spore cultures, 42 

Spot inoculation, 39 

Streak cultures, 41 

Three-point inoculation, 39, 40 
Currie, J.N. ,237, 328 
Curzi, M., 163, 166, 321 
Czapek's solution agar, 32 

D 

Dale, Elizabeth, 27, 101 , 321 
Dangeard, P. A., 27, 322, 28, 30 
De Bary, A., See Bary, A. De 
Descriptive sheet, 82 
Descriptive terms, 11 
Desiccation of molds, vacuum, 53 
Dewar flask, 53, 54 
Diastatic enzymes, 259, 257 
"Digestin", 213 
Dilution cultures, 40 
Dimargaris, 9 

Diploslephanus Langeron, 8 
Disjunctor, 25 
Dodge, B. O., Ill 
Dodge, C. W., 5, 154, 322 
Doelger and Prescott, 237 
Dox,A.W.,239,322,32 
Dried cultures — See Lyophil preserva- 
tion of molds 
Dried specimens, 62 
Drierite, 53 
Dual character of A. janus, 188, 189 



E 



Earle, F. R., 178 
Ecads, 64 



Eidam,E.,155,156,322 

"Eidamsche blasen", 28 

Elser, W. J., et al., 53, 322 

Emericella variecolor Berk, and Br., 9, 

163 
Emmons, C. W.,198 
Endogenous conidia, 25 
hmgler and Prantl, 6 
Enzymes, 238, 249, 213, 257, 270-271, 194, 

302-306 
Enzyme production 

by A . flavus-aryzae group, 270-271 , 302- 
304 

by A. niger group, 238, 304-305 

by A . okazakii, 213 

by A . tamarii group, 257 

by A . wentii group, 249 
Ergosterol, 194,317 
Escherichia coli, 154 
Euaspergillus Ludwig, 8 
Eurotiaceae, 6 
Eurotiales, 6 
Eurotium Link, 7, 101 
Eurotium herbariorum, 101, 125, 127 



Fat production, 194, 238, 306-307 
Ferdinandsen and Winge, 17, 228, 322 
Fermentation Division, N.R.R.L., ix, 

205 
Fernbach, A., 238, 304 
Fish, fermented, 286 
Fisher, Ed., 165 
Flavicidin, 272 
Flavicin, 272 
Flavipes group, 179-182 

Antibiosis, 182 

Color reactions, 180 

Occurrence, 182 

Outstanding characters, 179 
Flavus-oryzae group, 259-272, 69, 68 

Antibiosis, 271-272 

Enzyme production, 270-271 

Kojic acid, 270 

"Moldy bran", 271 

Occurrence, 269 

Outstanding characters, 259 

Pathogenesis, 271 

Variation in, 68-69, 260 
da Fonseca, Olympio, 227, 230, 322 



368 



INDEX 



Foot-cell, 17, 19 

of A. effusus, 264 
Frazer and Chambers, 27, 322 
Fresenius, G., vii, 4, 275, 148 
Fumaric acid, 237, 293-294 
Fumigacin, 154 
Fumigatin, 154 
Fumigatus group, 148-154 

Allergy, 154 

Antibiosis, 154 

Economic importance, 153 

Occurrence, 153 

Outstanding characters, 148 

Pathogenicity, 154 

Thermophilic habit, 153 
Fungi Imperfecti, 6 

G 

Gallic acid, 238, 294 
Galloway, L.D., 75, 322 
Gene mutation, 63 
General bibliography, 319-330 
Generic diagnosis, 6 
Gilman and Abbott, 202, 322 
Glaucic acid, 299 
Glaucus group, 100-147 

Economic importance, 146 

Group relationships, 102 

Historical considerations, 101 

Laboratory cultivation, 101 

Occurrence, 146 

Outstanding characters, 100 

Pathogenicity, 146 

Pigment formation, 15 

Temperature relations of, 45, 133, 134 

Variation in, 143-145 
Glister, G. A., 272, 300 
Gluconic acid, 238, 295-297 
d-Gluconic acid, 299 
Glucuronic acid, 299 
Glycolic acid, 299 
Glyoxylic acid, 299 
Gould, B.S., 258, 298 
Gould and Raistrick, 15, 323 
Graphic key to groups, 87 
Greene, H. C, 65, 323 
Greene and Fred, 56, 323 
Group keys 

A. candidus group, 207 

A. clavatus group, 92 



A.flavipes group, 179 

A . flavus-oryzae group, 259-260 

A. fumigatus group, 148 

A. glaucus group, 102 

A. nidulans group, 155-156 

A. niger group, 215-216 

A. niger group (Mosseray's), 232-235 

A. ochraceus group, 274-275 

A. tamarii group, 250-251 

A . terreus group, 195 

A. ustus group, 171 

A. versicolor group, 183 

A. wentii group, 241 
Growth substances, 316 
Guegen, F.,4, 323 
Gum, 317 

H 

Haines, R. W., ix, Plates I-VII 

Haller, D. A.,3 

Hanging drop culture, 45 

Hansen, H. N., 65, 323 

Hansen and Smith, 65, 323 

Hanzawa, J., ix, 225 

Hao,L. C.,271,303,304 

Hay-infusion agar, 35 

Head, 16, 18-21 

Henrard, P., 27, 323 

Herbarium specimens, 62 

Herrick, H. T., ix, 237, 238, 227 

Heterothallism, 27 

High sugar Czapek's agar, 101 

Historical introduction, 3 

Hollaender, A.,75, 326 

Homothallism, 27 

Hooper, et al., 99, 323 

Huber,G.A.,278,286,323 

Hulle cells, 28 

in A. carneus, 176, 202 

in A . flavipes group, 179 

in A. nidulans group, 157, 167 

in A. ustus group, 176 

in A. versicolor group, 188, 192 

Hydrogen-ion concentration, 14, 34, 35, 
95 

Hydroxylamine, 317 

Hyphomycetes, 6 

I 
Identification of Aspergilli, 81-91 
Incubation of stock cultures, 51 



INDEX 



369 



Incubators, 48 

Induced variation, 74 

Industrial Farm Products Research 

Division, 227 
Inoculating needles and loops, 47 
Intermediate species, 86 
Interpretation of Descriptions, 10 
Inui, T., 230 
Invertase deficiency in A. panamensis, 

244 
Inzengea erythrospora Borzi, 9 
Isolation of single spores, 43, 44 
Itaconic acid, 204-205, 143, 297 



Jardin Botanique de l'Etat, 235 
Jones, Rake, and Hamre, 272, 301 

K 

Karow,E.O.,249,293,323 
"Katsuobushi", 286 
Keitt, G. W., 43, 323 
Keys, 86-91 

based on color, 88-89 

based on morphology, 90-91 

Graphic, 87 

to groups, 86-91 

to species — see group keys 
Kinoshita, K., 142, 143, 323, 324 
Kita,G.,ix,255,260,324 
Kluyver and Perquin, 270, 298 
Koji,38,249,257,260 
Kojic acid, 249, 258, 270, 297-298 



Lacto-phenol, 48 
Lambert, E. B., 43, 324 
Langeron,M.,204, 324 
La Rue, CD., 43, 324 
Ledingham, G. A., ix, 198 
Link, H. F.,3, 101,210 
Linossier, G., 206, 324, 15, 236 
Lockwood, et al., 76, 324, 205, 297 
Loops, transfer, 47 
Ludwig, F.,8,324 
Lutz,L.,226 

Lyophil preservation of bacteria, 53 
Lyophil preservation of molds, 53 
Advantages of, 56 



Apparatus, 54 
Methods, 53, 55 
Recultivation, 55 
Viability, 53 

M 

Ma, Roberta, ix 

Macroaspergilli, 16 

Macy, H., ix 

Maintenance of cultures, 51-57 

Malic acid, 299 

Malt extract agar, 35 

Mangin, M. L., 118, 122, 127, 143 

Mannitol, 317 

Manual, use of, 81-91 

Marchal,E.,253,324 

Martin, G.W., 6 

Mary Clare, Sister, 285 

May, O. E., ix, 227, 237, 238, 270 

McCoy, Prof. Elizabeth, 56 

McKee and MacPhillamy, 272, 301 

McKee, Rake, and Houck, 272, 301 

Mealy bugs, 267, 260 

Media, culture, 31 

Czapek solution agar, 32 

Hay infusion agar, 35 

High-sugar Czapek agar, 101 

Influence of, 36 

Malt extract agar, 35 

Mosseray's Raulin's solution, 34 

Neutral Raulin's solution, 33 

Sporulation, 37, 38 

Steep-liquor Czapek's agar, 35 

Steinberg's solution, 34 
Melleic acid, 299 
Metarrhizium, 239 
Methyl cellosolve, 53 
Micheli, P. A., vii, 3, 100 
Microaspergilli, 16 

Microbiological Congress, Third Inter- 
national, 5 
Microscopes, 48 
Mildew, 146, 239 
Mites, 60 

Damage caused by, 61 

Elimination of, 61-62 

Poison, 61 
Mixed cultures, 57 
Mold Disease of A . niger, 59, 60 
Mold Tissue, Composition of, 301-302 



370 



INDEX 



"Moldy bran", 271 

Molliard,M.,238,295 

Monilia, 3 

Monograph, 5 

Monospore Cultures, 42 

"Monster", 74, 85 

Montagne, J. F., 3 

Morphology and Description, 10 

Morrow, Marie B., ix 

Mosseray, Raoul, ix, 70, 75, 83, 229, 232, 

235 
Mosseray 's Synopsis of Species, 232-234 
Mounting fluid, 47 

Moyer, A. J., 37, 227, 237, 238, 258, 270 
Moyer and Coghill, 205, 297 
M-type and C-type, 65 
Mucidinaceae, 6 
Mucidineae, 6 
Mucoraceae, 42 
Mucor herbariorum, 7 
Mutant, definition of, 63 
Mutation, 63 
Mutations, deficiency, 76 
Mutations, induced, 74 

by cathode rays, 75 

by chemicals, 74, 75 

by heat stimulation, 74 

by ultra-violet radiation, 75, 76, 77 

in A. glaucus group, 74 

in A. niger, 75 

in A. terreus, 75 
Mutations, "injury", 75 
Mutations, morphological, 76, 77 
Mutation, natural, 71 

in A . fumigatus, 72 

in A. glaucus group, 73 

in A. nidulans, 72 

in A. niger, 72, 73 
Mutations, physiological, 78 
Mutation, taxonomic usage, 84 
Nakazawa, R., ix, 225 
Nakazawa, Simo, and Watanabe, 220 

N 

Natural groups, 82, 87 

Natural Mutation, 71 

Natural relationship of groups, 87 

Needles, transfer, 47 

Neill, J. C, 5, 69, 325, 83, 143, 193 

Neutral Raulin's solution, 33 



New species, 84 

Bases for description, 85 

Recognition of, 84 
Nidulans group, 155-170 

Occurrence, 170 

Outstanding characters, 155-156 

Pathogenicity, 170 
Niger group, 214-240 

Citric Acid, 237, 290-293 

Coloration, 236 

Conidiophore structure, 19, 222 

Enzyme production, 238, 304-305 

Fat production, 238, 306 

Fumaric Acid, 237, 293-294 

Gallic Acid, 237, 294 

Gluconic Acid, 238, 295-297 

Industrial strains, preservation, 240, 
50-62 

Mildew, 239 

Mosseray 's Synopsis of Species, ^32- 
234 

Occurrence, 236 

Outstanding characters, 214 

Oxalic acid, 238, 298-299 

Pathogenesis, 239-240, 307-310 

Physiology, 239, 310-312 

Soil analysis, 238, 313-314 

Strain "67", 227 

Variation in head structure, 218 

Variation in spore size, 221 
Niklas, H., 238, 313 

Northern Regional Research Laboratory 
vii,50 

O 

Ochraceus group, 273-286 

Fermentations, 286 

Occurrence, 285-286 

Outstanding characters, 273 
Ochracin, 318 
Odor 

Actinomyces-like in A. foetidus, 219 

foetid in A. clavatus, 95 

foetid in A. flavipes, 180 
Okazaki,K., 213, 211,325 
Oryzae group — See Flavus-oryzae group 
Oshima, K., ix, 260, 303, 261, 270 
Ota, M., 271,308 
Overgrowths, 58 
Oxalic acid, 238, 298-299 
Oxford and Raistrick, 154, 325 



INDEX 



371 



Parasiticin, 272 
Parasitism of Aspergilli, 59 
Partansky and McPherson, 239, 325 
Pathogenicity, 307-310 

in A . flavus-oryzae group, 271 

in A. fumigatus group, 154 

in .4 . glaucus group, 146-147 

in A. nidulans group, 169-170 

in .4. niger group, 239-240 

in A. terreus group, 204 

in A. versicolor group, 193-194 
Patouillard and Delacroix, 222, 223 
Patulin,98 
Penicillin, 99, 182 
Penicillin-like substances 

in A. flavipes, 182 

in A.flavus, 272 

in A. giganteus, 66 
Penicillium chrysogenum Thorn, 57 
Penicillium notatum Westling, 272 
Penicillium patulum West., 99 
Penicillium restrictum Abbott, 67, 186 
Penicillium rugulosum Thorn, 59 
Penicillium spinulosum Thorn, 154, 300 
Perithecia, 26 

in A . fischeri, 149, 151-153 

in A. glaucus group, 27, 100-137, 106, 
115 

in A. nidulans group, 155-166, 161, 164 
Persoon, C. H., 3 

Pfizer (Chas.) and Co. Inc., ix, Pis. I-VII 
Phialids, 23 
Philpot, F. J.,99,301 
Photographic equipment, 49 
Physiology, 239, 310-312 
Pigments, 15, 143, 236, 312-313 
Plectascineae, 6 
Plug poison, 61 
Poison, cotton plugs, 61 
Polysaccharides, 318 
Pontillon, C, 238, 306 
Preservation of Cultures. 51 

in agar slants, 51-52 

in lyophil form, 53-56, 52 

in soil, 56-57, 52 

on vegetable substrata, 57 

Industrial strains, 240, 50-62 
Progressive variation, 68 



Proteolytic enzyme, 213, 257, 271 
Pulli, 214 

Q 

Quilico and Di Capua, 15, 313 

R 

Raistrick, Harold, ix, 111, 117, 143, 231, 

238, 283 
Raistrick, Robinson and Todd, 15, 313 
Rami, 23 
Raper, Coghill, and Hollaender, 75, 326, 

205, 206, 212, 225 
Raper and Thorn, 166, 175, 187, 190, 242, 

283 
Raulin, J.,4,237 
"Raulin neutre-gelose", 70, 33 
Raulin's solution, 34 
Recultivation of dried cultures, 55 
Reduced structures in A. sydowi, 184 
Rejected species — See Check list of 

species 
Resting nuclei, 73 
Ridgway, Robert, 13, 326 
Ridley, H. N., 13,64,326 

S 

Saccardo, P. A., 11, 165, 326 

Saccharification, 271 

Saito,K.,231,297 

Salmonella typhi-murinum, 154 

Saltant, 64 

Sartory , A . , 4 

Sartory and Meyer, 212, 327 

Sartorya fumigata , 9, 153 

Scales, F. M., 253 

Schiemann, Elizabeth, 74, 223, 224, 327, 

206 
Schmidt, C.F., Jr., 238, 306 
Schwartz, W., 30, 327 
Sclerotia, 30, 212 

in A. candidus group, 212, 208 

in A. flavus-oryzae group, 259, 265 

in A. niger group, 214, 217 

in A. ochraceus group, 273, 277 

in A . tamarii group, 250 

in A . wentii group, 241 , 243 

significance of, 212 

structure of, 30 
Secondary growth, 58 



372 



INDEX 



Sectors, 64 

Septa, 17 

Shih, Y. K.,ix,198 

Short-lived species, 51 

Siebenmann, F. , 271 , 307 

Simonart, Paul, ix, 232 

Single-spore cultures, 42, 44 

Smear cultures, 41 

Smith, George, ix, 5, 117, 102, 137, 139, 

141,246,285 
Sodium lauryl sulfonate, 37 
Soil analysis, 238, 313-314 
Soil Fungi, Summary of, 202 
Soil Preservation of Molds, 56 

Directions, for, 56 

Viability, 56 
Sopp,0. J.O.,9 
Soy products, including soya sauce, 249, 

258, 270 
Speare,A.T.,260,266,328 
Species 

Accepted, 360-361 

Check list of, viii, 331-359 

Definition of, 83 

Diagnosis of, 82 

Keys to, see group keys 

New, 84 

Rejected — See Check list of species 

Transitional, 86 
Specimens, identification from, 81 
Spegazzini,C.,8,229,328 
Spinulosin, 154 
Spores 

Ascospores, 28, 29, 152, 162, 130, 108 

Conidia, 23-25, 24, 221 
Sporodinia, 3 
Sporulation media, 37, 38 
Sprays, 62 

Stalk — See Conidiophore, 17 
Staphylococcus, 53, 98, 99 
Staphylococcus aureus, 154, 182 
Steep liquor Czapek's agar, 35 
Steinberg, R. A., 239, 310, 311, 312 
Steinberg's nutrient solution, 34 
Steinberg and Thorn, 75, 328, 206, 2^2, 

224, 225, 240 
Sterigmata, 22 

Primary, 19, 23 

Secondary, 19,23 
Sterigmatocystis Cramer, 8 
"Sterile hyphae" in A. unguis, 169 



Stock cultures, 51-57 
Agar slants, 51 
Cultivation, 51 
Incubation, 51 
Periodic transfer, 51 
Storage, 51 

Streak cultures, 41 

Streptococcus viridans, 154 

Substrata, see Media 

Swift, MarjorieE., Ill 

Synonyms 

in A. candidus group, 211 

in A. chevalieri series, 120, 121 

in A. clavatus group, 98 

in A. echinulatus, 133 

in A. flavus-oryzae group, 266, 269 

in A. luchuensis series, 232 

in A. niger series, 225-226 

in A. niveo-glaucus , 137 

in A. ochraceus series, 281 

in A. repens series, 107, 110 

in A. restrictus series, 139 

in A . ruber series, 117 

in A. sulphureus series, 276 

in A. sydoivi series, 186 

in A. tamarii group, 256-257 

in A. terreus group, 197, 200 

in A. umbrosus series, 131 

in A. versicolor series, 192, 193 

in A. wentii group, 247-248 

Synonymy, 6 

"System und Phylogenie", 5 



Takahashi,T.,260,328 
Takamine, J., 270, 302 
"Tamari", 258 
Tamarii group, 250-258 

Economic importance, 257-258 

Occurrence, 257 

Outstanding characters, 250 
Tamiya and Morita, 4 
Tannic acid, 237, 294 
Tannin, 237, 294 
Taubenhaus, J. J., ix 
Temperature, 45, 46 
Temperature, effect of, 

on A. giganteus, 45, 97 

in A. glaucus group, 45 

on A . janus, 45, 187 

on A. medius, 45, 46, 134 



INDEX 



373 



Temperatures, optimum, 45 
Terminology, descriptive, 11 
Terrein, 318 
Terreus group, 195-205 

Antibiotics, 205 

Itaconic acid, 204, 205, 297 

Mutations, 75 

Occurrence, 204 

Outstanding characters, 195 

Pathogenesis, 204 

Variation in, 66, 67, 197 
Tessar lenses, 49 
Thaxter, R. T., 50, 189, 251 
Thermophilic species, 45 
Thorn, C, vii,5, 195,328 
Thorn and Church, vii, 4, 5, 102, 137, 206, 

235, 255, 260 
Thorn and Raper, 74, 328, 102, 168 
Thorn and Steinberg, 75, 328, 144, 150, 

212,221 
van Tieghem, Ph., vii, 4, 237 
Timonin, M. I., 203, 205, 329, 213 
Topical bibliography, 289-318 
Transfer needles and loops, 47 
Transfer of stock cultures, 51 
Transitional species, 86 
Trichoderma, 42 
Turfltt,G.E.,246 
"Type" culture collections, 50 

U 

Ultra-violet radiation, 75, 76, 77 
Underkofler et al., 271, 303, 285 
Ustilago phoenicis Cda., 223 
Ustus group, 171-178 

Hulle cells, types, 176 

Occurrence, 178 

Outstanding characters, 171 

V 

Vacuum desiccation of molds, 53 

Vacuum tester, 55 

Variant, 64 

Variation, natural, 63-78 

in A. fischeri, 65 

in A . flavus-oryzae group, 68, 69 

in A . fumigatus, 68 

in A. niger group, 70, 71 

in A . sydowi, 67 

in A. terreus group, 66, 67 

Intra-group, 68 



Intra-species, 66, 67 

Intra-strain, 65 
Variation, references, 314-315 
Variety, taxonomic usage, 84 
Vegetable substrata, 57 
Versicolor group, 183-194 

Occurrence, 194 

Outstanding characters, 183 

Pathogenesis, 193 

Strain variation, 192, 186 
Vesicle, 18, 19, 22 
Vestigial characters, 82 
Vibrio cholorae, 154 
Vitamin D, 238 
Vitamins, 316 
Vuillemin, P.,165,329 

W 

Waksman and Bugie, 272, 301 

Waksman et al., 98, 300, 99, 301, 154 

Ward, G.E., 237, 270, 329 

Webb, P. H.W.,97,329 

Wehmer, C, vii, 4, 226, 237, 238, 247, 283 

Weisner, B. P., 98, 300 

Wells, May, Moyer, Herrick, et al., 227, 

237, 270 
Wentii group, 241-249 

Economic importance, 249 

Occurrence, 249 

Outstanding characters, 241 
Westerdijk, Johanna, viii, 110, 142, 283 
Whelden, R. M., 75, 330, 206, 225 
White; E. C, 182, 330, 271, 300, 272 
White mutants, 206 
Wickerham, L. J., 55 
Wickerham and Andreasen, 53, 330 
Wiggers, Fredricus Henricus, 7, 100 
Wilhelm, K. A., vii, 4, 281, 330 
Wilkins and Harris, 99, 300 
Wire, nichrome, 47 

platinum-iridium, 47 
Wolf, F. A., 97, 330 

Y 

Yabuta, T., 249, 297, 330 
Yuill, Edward, 65, 150, 159, 206, 212, 221 
Yuill, John and Edward, ix, 73, 145, 240 
Yukawa, M., 279, 286, 330 

Z 

Zea Mays, 267 
Zonation, 11-13, 12